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Single-cell genomic sequencing using Multiple Displacement Amplification.pdf

Single-cell genomic sequencing using Multiple Displacement Amplification.pdf

ID：34142164

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页数：7页

时间：2019-03-03

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1、Single-cellgenomicsequencingusingMultipleDisplacementAmplificationRogerSLaskenSinglemicrobialcellscannowbesequencedusingDNAwithoutrequiringdevelopmentofculturemethods.amplifiedbytheMultipleDisplacementAmplification(MDA)WhileamplificationbiasintheMDAreactioncanresultreaction.ThefewfemtogramsofDNAinabact

2、eriumareinunderrepresentationandlossofsomesequences,theamplifiedintomicrogramsofhighmolecularweightDNArewardsaregreatoftenprovidingthefirstgenomicsuitableforDNAlibraryconstructionandSangersequencing.sequencesobtainedfromnovelspecies.Furthermore,TheMDA-generatedDNAalsoperformswellwhenusedseveralstudie

3、shavealreadyachievedbetween50anddirectlyastemplateforpyrosequencingbythe454Life75%completionofgenomesfromunculturedsinglecells,Sciencesmethod.WhileMDAfromsinglecellslosessomeofandmethodimprovementsarepromisingtoimprovethegenomicsequence,thisapproachwillgreatlyaccelerateuponthisfurther.Recentworkhas

4、alsodemonstratedthepaceofsequencingfromunculturedmicrobes.Thethepowerofsingle-cellsequencingtorevealthepresencegeneticallylinkedsequencesfromsinglecellsarealsoaofgenespredictedfromenvironmentalandphysiologicalpowerfultooltobeusedinguidinggenomicassemblyofstudiesevenfrompartialdraftgenomes.Thegenomi

5、cshotgunsequencesofmultipleorganismsfromenvironmentaldatacouldalsoassistintheprocessofeventuallydevel-DNAextracts(metagenomicsequences).opingculturemethods,stillaparamountgoalrequiredtofullyunderstandthebiologyoforganisms.Theidentifi-AddressesJ.CraigVenterInstitute,11149N.TorreyPinesRd.,Suite200,cat

6、ionofmetabolicpathwayscaninformthedesignofLaJolla,CA92037,UnitedStatesculturemediaandgrowthconditions.AnotheradvantageofMDA-basedsingle-cellgenomicsisthatitiseasilyCorrespondingauthor:Lasken,RogerS(rlasken@jcvi.org)automatedandcanbecombinedwithhighthroughputcellsortingoflargenumbersofmicrobes.PCRsc

7、reeningCurrentOpinioninMicrobiology2007,10:510–516oftheMDAreactionsforinformativesequences,suchasthe16SrRNAandrecAgenes,canbeusedtoidentifytheThisreviewcomesfromathemedissueongenomesofinterestforadditionala

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