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[Music]

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[Applause]

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thank you all one quick comment

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there are even shorter what tri cycles

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than Days and seasons Bruce and I

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visited volcanic hydrothermal fields all

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over the world really

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and there are wet/dry cycles measured in

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minutes to an hour or so because of hot

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spring activity fluctuating levels of

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water that just suddenly keep in mind

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we're not stuck with really long daily

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or seasonal wet/dry cycles okay I gotta

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give you a few minutes of introduction

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to get to the unpublished material I

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want to talk to you about reading button

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okay okay so a lot of what I'm showing

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you is in fact the experimental results

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and that leads to a hot spring

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hypothesis that is testable the two

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examples of hydrothermal regions that

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have been suggested in the literature by

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ourselves and by the people interested

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in hydrothermal events is the so-called

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black smokers and alkaline vents shown

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on the Left which in fact are saltwater

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and the hydrothermal fields that we're

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talking about which is distilled

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freshwater distilled by evaporation from

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a global ocean and falling on volcanic

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land masses I only have time to talk to

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you about the freshwater so the

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favorable properties that we've observed

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in French water is that organic

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compounds accumulate on land masses as

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opposed to being diluted into the ocean

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and as Ben had just suggested they go

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through a cyclic process and the the

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deposition is a constant rate we have

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found that these pools tend to be acidic

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which we think is important for the kind

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of chemistry we're doing and the acidity

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results from so2 that is a volcanic gas

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it turns into so3 a weak acid

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the pools that we worked with are all

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down around pH 2 to ph 3 on the volcanic

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areas we've been testing the

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concentration of ions is very low Mille

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molar concentration wet/dry cycles

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concentrate potential reactants and

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those condensation reactions can then

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occur in the low water activity and

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we've done a thermodynamic analysis of

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this and it's very clear that there's

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enough chemical energy produced by the

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wet/dry cycle to drive ester bond

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synthesis finally the elevated

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temperature up in the 80 to 90 degree

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range provides activation energy for the

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formation of phosphodiester linkages and

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last of all if there are lipid like

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components present the polymers get

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encapsulated to become what we call

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protocells these are steps toward life

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they're not life itself but they are

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certainly able to evolve and that's

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where we are now in our research so we

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wanted to test this hypothesis we

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fabricated a simulation chamber that

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puts small vog laz files through what

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dry cycles we control carbon dioxide

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elevated temperature acidic pH range and

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the cycle is what we've talked about so

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the the device that we've built up

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depends on this relatively low energy

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synthesis of ester bonds and I want to

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make this clear to you if you mix

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ethanol and acetic acid there's a

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spontaneous reaction in which water

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comes off and ethyl acetate is produced

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ethyl acetate is the ester that is part

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of nail polish remover if you were

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probably all familiar with that smell

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however the same kind of reaction can

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occur between phosphate and the O H

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groups on a couple of nucleotides such

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as adenosine monophosphate shown here

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and if we want to pull the reaction to

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the right all we have to do is have a

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way for water to leave the reaction so

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there's no back reaction and in both

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cases we should expect to see

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Station reactions leading to fossil

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ester linkages in the bottom thing here

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we have the device that we built up but

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it's got a source of carbon dioxide

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basically keeping out air air and oxygen

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doesn't turn out to be terribly

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important to us but we just have co2 as

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a way to reduce the possibility of

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oxidation reactions there's a heat

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source that brings up that aluminum disk

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to about 80 to 90 degrees our choice of

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temperature and that disk rotates about

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once an hour and brings the vials under

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a gas flow of carbon dioxide which tries

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them down and then down at the bottom

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right you can see a hydration cycle this

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is a syringe pump and the water is

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dripping into the vials at those two

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points so at the end of a number of

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cycles we will pull the stuff out and

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analyze it so I'm going to show you one

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that we've done with a MP and UMP

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mixture we can have a lipid matrix there

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to promote the reaction we found that

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it's favorable and has a protective

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effect on polymers we do multiple

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wet/dry cycles we bring down the

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products with ethanol precipitation a

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very standard way to bring down polymer

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products we do that by spin tube or

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ethanol precipitation then we run either

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gel electrophoresis or we run nanopore

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analysis and you're going to see a

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little bit of results of that so the

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first thing we did ten years ago now

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2008 with Souter Rajamani and in the lab

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was to see whether these polymers could

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be in labelled with enzymes that

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recognize biological RNA if we've made

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anything that seems that looks like a

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biological RNA these enzymes should at

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least label the ends of those molecules

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and it's the in labeling southland

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phosphatase t4 kinase radioactive ATP

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you end up with a radioactive phosphor

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RNA labeled with radioactive phosphate

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so this is what su

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put together each of those lanes these

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are gel electrophoresis lanes are

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separate experiments it works again and

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again and again it's very robust all of

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these are different conditions that we

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put it through I just want to point out

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two of these this is the number of

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cycles we get more and more and more as

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we increase the number of cycles and

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look at the length of these polymers

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compared to this ladder here this is a

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RNA ladder with known RNA links and

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right next to it you can see that these

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polymers range from 10 burrs up to over

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hundred Murs in length that's really

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quite extraordinary that's spontaneously

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we can make polymers long enough to be

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labeled by these enzymes of recognize a

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MP I don't have time to show you all of

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those but just a variety of conditions

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or different lipids and different ratios

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and so forth so you can also see these

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by you know also see these by

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fluorescent labeling where we have a dye

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that binds to a polymer such as RNA here

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we have two three kinds of RNA that are

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known RNA this is a home polymer called

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poly and Milic acid there's another home

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polymer or that's part of your daily

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poly Abney leak acid poly a notice that

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they die which is Atheneum and a Curie

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intercalating guy does not strongly

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label these single numbers but if we

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make a mixture where we get in fact a

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double helical form of a poly a poly

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duplex structure now these micro gram

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quantities are labeled by the dye

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because the dye intercalates between the

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stack bases and becomes fluorescent and

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here is our polymer product again

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ranging from 20 immers up to the Han

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River range

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here's another dye called cyber safe and

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here's our product here's a deist double

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stranded 20 mer right there and here's

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our product so we were pretty sure that

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we were making something that was a

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polymer but we wanted to try one more

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way to look at this so we did what are

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what is

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called nanopore sequencing now this is

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new so this isn't been published yet

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we're repeating this so these are sort

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of preliminary work I'm going to show

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you now so the idea of nanopore

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sequencing is that we can lead a double

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strand of DNA or RNA in fact into a

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molecular motor which ratchets the DNA

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basis at about 450 nucleotides per

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second through a limiting aperture in a

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protein Nana pour that aperture has

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about three bases in it at any given

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time as the bases are ratcheted through

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that area we have a change in the

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electrical current in the Pico ampere

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range which reflects the base sequence

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so this is now out as a commercial

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device that's called the min ion I'm

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going to show you one of those and also

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the Promethean this is Oxford nanopore

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technology building on the nanopore idea

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so let's take a look at the next slide

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here is what I'm going to show you we

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wanted to be sure that we really could

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make something that was exactly the same

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as a strand of DNA so we kept it simple

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we decided to use TMP thymidine mono

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phosphate as the monomer this can only

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make 3-5 fossil ester linkages we put it

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through multiple wet/dry cycles we

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expect to get all ago Simon Dilek acid

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just a whole bunch of tees in a row and

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then we can isolate that we like a tit

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to a known strand of DNA in order to

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differentiate between the signal we're

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going to get from the ala goatee and a

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known base sequence of DNA and then we

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put that through nanopore sequencing so

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here's what it looks like to use the

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minion your sample is injected into this

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handheld device it goes to 2000 nana

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pours in this little area here that's

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the sensor chip each one is

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independently being being referenced by

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the electronics of the system all of the

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electronics are are in this device and

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the signal is fed into your laptop

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computer this is gone all over the world

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now looking for Ebola in Africa every 90

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minutes it goes over our heads and then

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in the international space station

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the midnight is being used sequenced the

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DNA of the organisms that now coat the

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interior of the ISS so let's take a look

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now is what we see what we see as a

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single nucleus in gold nucleic acid

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going through the pore looks like this

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this is the known sequence going through

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and they'll up and down is in fact the a

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G C T of that known bit of DNA and then

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right there is this long string which is

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just the what we think is the Aldo

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goatee homo polymer passing through the

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pore we have a base calling device that

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allows us to turn this electrical signal

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into the nucleobases

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and this is what it looks like here's a

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gel of the radioactively-labeled i'll

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ago simon teens starting down around 30

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Murs going all the way up to about 90

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Merce you can see these individual all

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ago Simon teens showing up in this gel

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and here are the base called fragments

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the ala goatee we see again and again

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attached to the known RNA ligated to the

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no night and this then is the alla

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goatee now attach that we have

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synthesized this at least convinces us

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that we can make very long strands of

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nucleic acids using just wet/dry cycles

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the 90 the 30 mer by the way is down in

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this range you can see that these links

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match so we see here the this is an

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intermediate length approximately v 40

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mer range then here is a very long one

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up in the ATM array

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okay now what we want to do is to see

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whether this actually works in out in

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the wild

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well I won't show you this - this stuff

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accumulates inside the lipid vesicles

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who we have a little bit there we can

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stain it with faculty in orange this is

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an are starting with a and P and um P

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these are the lipids and the

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encapsulated material after about three

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cycles ear is with DNA the this is a da

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MP and C and sorry TMP and you can see

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the Dappy now staining these vesicles

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with this encapsulated DNA so we're

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pretty sure that not only can we make it

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but it becomes encapsulated to make

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protocells now we're going to take out

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in the wild and ask this last question

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in my talk does it actually work in

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conditions such as we assumed were

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available on the early Earth what you

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just saw is a hot spring in Rotorua

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Bruce Bruce saw was down there just last

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summer doing this work so he's my

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co-author on this talk he would put

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these tubes in a large aluminum plate

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there's approximately a hundred of them

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they have the dried material that I did

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in the lab at UC Santa Cruz and he put

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these through four cycles about 3045

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minutes each and he added water from an

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acidic hot spring for the recycling so

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four cycles here is the products they

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were getting out notice that the

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products containing you were more

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abundant than the products containing a

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which was interesting we didn't expect

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that but that's just the way it turned

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out we then did a gel of this and

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compared what I showed you in that first

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gel with the products here's what I

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showed you the first time around and

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here is what we see in the hot spring

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cycles so we're convinced that this is

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not just the laboratory phenomenon but

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also can

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or out there in the real world and I

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recommend that people are doing work in

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the laboratory once in a while

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step out of the lab into a place that we

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consider be a prebiotic analog

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environment and see if your result

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actually works under real conditions so

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finally I'll end up with our conclusions

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this is an alternative scenario that

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we're developing for the origin of

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cellular life we think that life began

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in freshwater hydro thermal pools

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subject to cycles life did not invent

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nucleic acids instead life discovered

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pre-existing hydrothermally cycled

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polymers and encapsulated them to make

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protocells the if amphiphiles are

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present they get encapsulated if you

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have proto cell populations they vary

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just as is always required for evolution

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selection and evolution there had to be

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variables they can undergo selection and

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evolution and what we're now looking for

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is Dutch how to test this conjecture the

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life began when rare systems of

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encapsulated polymers happen to be

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capable of the catalytic functions

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related to the light process we put this

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into a sort of a geological scenario and

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that's just my last slide now and we

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have a synthesis of organic material

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being accumulated on these volcanic land

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masses the as these pilot of these pools

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go through cycles and the sugar gets

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concentrated it begins to react here's

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the reaction cycle you can see going

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around this circle between a wet phase

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down drying to a dry phase and back to a

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wet face this cycle then builds up cycle

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after cycle increasingly complex

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protocells as they accumulate they get

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distributed downhill toward the sea

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water ocean and finally adapt to sea

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water where they colonize the ocean as

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the last step in this origin of life

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scenario

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thank you I'm afraid we're a little bit

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limited for time but please find dr.

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Deemer if you have any questions for him

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and we'll go to our next speaker Vincent