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  • Illumina Sequencing Train Online

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    illumina sequencing train online

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    131 Ansichten3 Seiten

    Illumina Sequencing Train Online

    Hochgeladen von

    api-274105371
    Illumina sequencing Train online this website uses cookies. By continuing to browse this site, you are agreeing to the use of our site cookies. To find out more, see our Terms of Use.

    Copyright:

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    von 3

    10/29/2015

    Illuminasequencing|Trainonline
    Thiswebsiteusescookies.Bycontinuingtobrowsethissite,youareagreeingtotheuseofoursitecookies.To
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    Trainonline
    trainingonlinecourselistebinextgenerationsequencingpracticalcoursewhatnextgenerationdnasequencingillumina

    Illuminasequencing
    InNGS,vastnumbersofshortreadsaresequencedinasinglestroke.
    Todothis,firstlytheinputsamplemustbecleavedintoshortsections.Thelengthof
    thesesectionswilldependontheparticularsequencingmachineryused.
    InIlluminasequencing,100150bpreadsareused.Somewhatlongerfragments
    areligatedtogenericadaptorsandannealedtoaslideusingtheadaptors.PCRis
    carriedouttoamplifyeachread,creatingaspotwithmanycopiesofthesame
    read.Theyarethenseparatedintosinglestrandstobesequenced.
    TheslideisfloodedwithnucleotidesandDNApolymerase.Thesenucleotidesare
    fluorescentlylabelled,withthecolourcorrespondingtothebase.Theyalsohavea
    terminator,sothatonlyonebaseisaddedatatime.

    Animageistakenoftheslide.Ineachreadlocation,therewillbeafluorescent
    signalindicatingthebasethathasbeenadded.

    https://www.ebi.ac.uk/training/online/course/ebinextgenerationsequencingpracticalcourse/whatnextgenerationdnasequencing/illumina

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    10/29/2015

    Illuminasequencing|Trainonline

    Theslideisthenpreparedforthenextcycle.Theterminatorsareremoved,allowing
    thenextbasetobeadded,andthefluorescentsignalisremoved,preventingthe
    signalfromcontaminatingthenextimage.
    Theprocessisrepeated,addingonenucleotideatatimeandimaginginbetween.

    Computersarethenusedtodetectthebaseateachsiteineachimageandtheseare
    usedtoconstructasequence.

    Allofthesequencereadswillbethesamelength,asthereadlengthdependsonthe
    numberofcyclescarriedout.

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