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[Music]

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[Applause]

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I'm delighted to be here today as a

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representative a team that's developed

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this origins of life laboratory in

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McMaster University the PI for this

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effort was Michael Reince Tedder

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biophysicist I am an astrophysicist

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young Fuli is a biochemist kind of a

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dream team in some way for an

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astrobiology setup I want to emphasize

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then that the key question that were

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interested in the origins of life one of

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the big questions is the role of

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information where does it come from how

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does it develop can it evolve the RNA

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world hypothesis provides a basis for

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thinking about that seems to be active

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in all young cells and coming to the

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talk earlier that we saw today the kind

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of environmental problems that you face

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in building up RNA polymers as you can

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see in warm little pond environments

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you've got UV that can destroy you

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you've got seepage at a bottom of ponds

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you have hydrolysis fighting you against

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this you've got a polymerization

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occurring at some rate in some kind of

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media and this is a beautiful

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experimental question that kind of

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defies easy simple theoretical address

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questions - so this has been our

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motivation in building up a laboratory

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which starts given that we're supposed

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to have molecules etc around nucleotides

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in this case lipids perhaps delivered

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from meteorites and the question is what

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happens next the first step how do we

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get polymers so in the as is shown in

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your diagram here we all know the

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importance of these condensation

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reactions particularly driven home by

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the work of David Deamer and his

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collaborators and in the situation that

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we're interested in warm little ponds or

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environments like this where we can

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easily have such warm wet dry cycles as

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well as thermal cycles that are driving

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the condensation reactions you see here

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some one of some nucleotides laying down

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on some layer which can be complicated

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there can be lipids clays salts various

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kinds of things mixed in this messy

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prebiotic environment we have heat and

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rise driving the thermal aspects of this

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the wedding and drawing allows you to

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form in one cycle a dime

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and there's some ability of these

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molecules afterwards during the wet

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phase during the next dry phase dimer

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can make dimer if appropriate conditions

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you have a former and by this process

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the important role of surfaces here can

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actually help this process along so this

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is the perfect kind of physical and

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chemical setup that motivates our

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laboratory as you see here this is

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designed by us and built by angstrom

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engineering of top-level technology

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company near Kitchener Ontario near our

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University that wonderful fanciful you

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know life generating machine and the

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frogs there is their artistic

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presentation the apparatus what you have

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on the Left pictures is Renet one of our

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students working in this it's a computer

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controlled situation we've got a

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temperature control we have control of

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the wet/dry cycles very carefully built

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in I'll show you in a moment

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we have pressures just going to one bar

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we can put different gasses into this we

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have a radiation field which is I can

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get my light going here the radiation

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field and eight and lamps carefully

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design LEDs from going from infrared to

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UV in these cylinder heads here nicely

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cooled so we I will show you the

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conditions here so looking into our lab

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into the simulator we've got various

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kinds of radiation fields showing you

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here

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our intent is to do this experiment as

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you define the conditions on other

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planets other planets getting the

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Trappist one system here are two spectra

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the solar spectrum is an example the

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Trappist one star so we are now putting

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together the program that will program

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in these radiation fields everything can

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be oscillated against everything else

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while the radiation is well the water

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and while you have wetness that can

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screen you from UV so at that point the

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UV would be off when you dry UV becomes

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a problem for you so the UV field turns

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on so all of these can be cycled in any

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manner that you wish

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for the given in piped environment that

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you want I'll show you this movie here

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so we're looking in the chamber this is

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the beautiful fall over the talk this

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morning we don't actually have a wet

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pond we do this by deuce that appear and

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disappear to 85% humidity so if you look

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inside the chamber you see that dude

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developing on the right-hand side you

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see the relative humidity dry up the

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peak that do piers

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there's a slight difference between the

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plate and the background so there's a

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due developing disappeared this can be

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beautifully controlled by the expert

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design or engineers and so we can very

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nicely produce any kind of wet/dry cycle

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the thermal cycling that you want to

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program in it's taken us a few months to

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find the optimal procedure for this but

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before you you have a machine that works

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extremely well so the way we prepare our

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many ponds these are many ponds I put on

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cyclic silicon wafers once the meter

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squared you pipette on whatever

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ingredients you think should be in your

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pond in whatever concentrations that you

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want here as shown four of them and

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shown here pipe heading on the important

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things during what dry cycles sorry

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about this you see the in the top here

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there's many layers of vesicles that

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layer one on top of the other during

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course of many cycles these layers that

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can be up to hundreds of layers here one

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atop the other and inside of each day as

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you trap some monomers dimers etc that

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find one of their mobile between these

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membranes they find one other they link

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up during the drive phase when you wet

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these things break up into little bags

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eventually and eventually you can

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encapsulate things so I will show you

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just some of our first results here

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first of all microscopy just with pure

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lipids the typical one used in all

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experiments with the dcmp with just a

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mixture of sorry of here a MP and UMP

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put in here so for this kind of a

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uniform field looking down and lipid

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in early with God we one cycle takes

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about an hour to do so we can do you

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know in one week we can get good way

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through 70 cycles make the cycle as long

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as you wish and effectively a minute at

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a month a year you know that doesn't

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matter just a number of cycles it

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matters here you see the growth of

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rather large structures a hundred micron

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100 micrometer is shown in here

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intermediates in the number of cycles

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you have these largest structures here

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which are starting to break up into the

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smaller vesicle type rain structures

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shown in here the walls get thicker and

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into this I'll show you where the

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nucleotides are in a moment in the final

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phase we get now on wrasse quite a

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complicated structure as these larger

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ones are broken up you have these VC Col

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like structures now inside about five

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foot about 50 microns or so as David

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Deamer others have found in their

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experiments and if I show you now just

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we're going to use like autofluorescence

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now these are nucleotides will glow

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they're excited and by light coming in

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here and the rate of the like the images

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now that you see here are actually

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mission from the nucleotides looking at

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their distribution so I'll go from our

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first stage to the later stages the last

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stages sequentially so here we see this

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uniform kind of distribution early on

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we've got these large empty regions that

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are formed the green as well nucleotides

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they're still kind of spread out but

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they're finding their way being

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concentrated in those walls a little bit

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later your few more cycles on you see

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that these larger structures are

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breaking up most of the nucleotides are

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being concentrated now in the walls by

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the very process we've been talking

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about a little bit further on you now

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see this very broken up kind of

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structure and peering into those on the

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scale you see that nucleotides are

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encapsulated very nicely

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we have various physical assays

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techniques by driving at these analysis

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for this including Neutron and here

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x-ray diffraction experiments so we can

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run on the samples to determine what's

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is the structure of the layers and where

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are the monomers in these layers these

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can be followed up by molecular dynamics

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simulations so here's the basic geometry

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shown down here the x-ray machine in the

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laboratory diffracting off this bilayer

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as an example this is the kind of 2d

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diffraction pattern you get the Bragg

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Peaks which you can then explore here

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we've got the various distance scale

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between the molecules between the lipids

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there's the bilayer here and by doing

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the analysis with some computer analysis

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behind you here they the distribution

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here in this diagram with the intensity

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as a function of Q here this shows the

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typical scales the typical scale in your

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bilayers might be 4.9 angstroms

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disorganised nucleotides have a

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separation of about 4.6 entrance that

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accounts for this peak and the organized

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nucleotides separated by about the

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distance of involve a phosphodiester

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bond I should say are in this growing

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peak right down here now if you follow

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this with time you find this graph here

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which is a question number of cycles

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we've gone to about 70 in this

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experiment by the way all this work is

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still unpublished please is this the

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first steps but this is just new stuff

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for showing you for the first time first

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results of our lab but you see the

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growing organized nucleotide fraction

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reaching up to about 10% now after about

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simony cycles in these organized units

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this graph doesn't tell you yet that

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they are actual polymers of course but

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they're organized like polymers to

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follow that up one wants to do things

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like a gel electrophoresis as an example

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as David Deamer was just showing

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so here is Renee preparing the samples

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we have centrifuge cleaning things up

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getting ready put the samples now gel

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electrophoresis columns which are shown

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here so this is again another part of

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our laboratory laboratory by the way is

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full equipped for all the physics

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biophysics setups a biochemical assays

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the various instrumentation we've been

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showing you and here I can just show you

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is the gel electrophoresis experiment

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here are two ladders 100 base pairs just

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a standard ladder everybody uses and

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again these molecules are negatively

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charged the lightest molecules move the

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furthest down in the gel that's the way

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this technique works and here is the

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result

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so this is the the ladder I showed you

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hundred base pairs this these are spread

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out a little bit because of salt effects

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we think but you can see in this pretty

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well resolved we've gotten already to

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about fifty mirrors perhaps we will

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check that but this has been with a

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rapidity kind of things I'm very

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interested in what David's results were

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in his own experiments but they seem to

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be working just fine here will of course

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want to do base sequencing of this work

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and in the next experiment in next part

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of the experiment but for what this says

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now given the reliability that David is

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pointed to for understanding an

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interpreted gel electrophoresis data

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this is hopeful that we're actually

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demonstrating this so I would just like

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to conclude that so far in these

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experiments we've been able to

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demonstrate under conditions of warm

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little ponds where we've got thermal

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cycles taking up to 80 degrees and down

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allowing bonds to form and dry wet

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cycles which allow for this alternate

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mobility and nailing things down to

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allow the polymers to grow that we're

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very effective in growing polymers that

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probably matter

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these have been encapsulated within

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these these vesicles we don't know how

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much of this RNA polymerize ation may

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occur inside the vesicle yet that's the

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next thing the next part of the

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experiment to try to determine that

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ultimately of course a huge step ahead

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we do not know if any of this material

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is capable of transcribing itself and

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that is the giant leap that of course

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we're all interested in the step towards

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some kind of evolution but I hope that

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you find this we feel that this is not

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only robust way of examining physics and

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chemistry in an early Earth environment

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but we will soon be able to ask the

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quest answered the edge question are we

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alone

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by doing this say in a Trappist one or

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any other kind of world you might want

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to dial up we will simply dial up a star

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radiation field and be able to run these

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under a variety of conditions so with

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that I'd like to thank you very much for

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your attention thank thank you Ralph -

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first question here in the centre I

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tomorrow Faraci from SPC Paris I I

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wanted to know if you did

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perhaps some low angle x-ray scattering

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to see if there is any super molecular

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structures that have a larger land scale

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than and then single a stacking distance

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and the answer to that is that and you

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know there's a range of structures that

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you do find there's a there's quite a

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distribution that you saw but still

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fairly heat and this has been followed

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up by molecular dynamics simulations so

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we think we have a good feeling between

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micro dynamics and the data as to you

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know what levels of structure we should

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be seeing yeah yeah but I was just

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meaning that at that q you are sensible

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to the the distance between the nuclear

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bases but if you go at lower Q you can

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look at the distance between the

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different layers or various other

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organization of the of the strength of

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oligomer that you have yeah we do probe

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the distance between layers I showed you

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that number I believe up there and the

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separation between

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yeah we get all those distances pretty

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well and second which is the range of

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concentration you think you are

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exploring during dry worked in cycles

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yes I think these were molar types

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setups in these experiments so and we

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actually forgot to mention we have equal

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a one on one of a MP and UMP and the

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mixtures that we had here which are

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actually good for building larger

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polymers as is known I got a question

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over here

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yes Ralph I just want to comment that

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implicit in your talk is the use of base

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pairing monomers the ANP and the UMP are

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in fact the base pairing of RNA and we I

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think you have found that the if we try

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to make these monomers out of a single

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nucleotide either poly Ujala go use or

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Allah Koei's it doesn't work nearly as

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well as having both prison that would be

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able to form a duplex strand and not

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just a single strand and we're getting

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strong evidence if that's the case from

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some atomic force microscopy that we're

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doing where we can actually see these

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strands accumulating I want to make a

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surface that's fascinating thanks for

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that remark one more question

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hi I'm Melina Popovich a blue marble

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space and such as science have you run

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nan de natura ng gel or excuse me

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denaturing gels no it's it's taken I'm

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not a biochemist myself we rely on

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collaboration with dr. yang Fu's labs

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laboratory to help us out with that it's

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been quite a trick just to get as you

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know it's very tricky to get RNA to go

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through gels probably because they the

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nature of the molecule they tend to ball

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up etc so it's been quite a trick even

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to get that data that I showed you

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I'm just not sure the extent of the

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experiment done with other gels yet I

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think this is the first one we've got

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working after some months of effort so

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we're still in the baby steps