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2015-BMMB852D:AppliedBioinforma8cs

Week15,Lecture30

IstvánAlbert

BiochemistryandMolecularBiologyandBioinforma8csConsul8ngCenter

PennState

RNA-SeqconceptsAnRNA-Seqanalysishasthreeseparateyetequallyimportantsteps

1.  Iden8fy/assignreadstoloca8ons

2.  Es8ma8ngabundancespertranscript

3.  Comparingabundancesàdifferen8alexpression

Thereisenormousdisagreementabouthoweachthesestepsshouldbeperformedàhenceverylargenumberofop8onsThereareworkflowsthatdoallthree.Thereareworkflowsthatmixandmatchfrommethods.

Makingsenseofyourdata

Soyou’veiden8fiedthetranscriptsthatdifferen8allyexpressacrosscondi8ons.

Nowwhat?

Thelengthyprocessofmakingsenseofdata

Themostimportantaspect

Don’texpectthedatatobothtellyouwhattheques:onis

andalsoprovidetheanswerforitHaveaques8on/hypothesisàthedatacananswerit.

Makingsensetakeslongerthanrunningtoolson

IPathwayguide–freetolookbutthencostsmoney

ErmineJ

Genesetanalysis

Paralysisofchoice

•  Toomanychoices–toomanyparameters

•  Whattopick,amIright

•  I’veseenexcellentstudentsstrugglewiththis

Trustyourselfandmoveon.Finishingthejobishardest.

Bioinforma8cscanbestressful

Beingtheonlyonethatunderstandshowtoperformsomeanalyseshashighsandlows•  Youcanbepartoflotsofpapers•  EasiertomakeyourownsuccessLotsoftruthinMickWatson’samusingblogpos:•  Aguidetothelonely(pet)bioinforma8cian

Genomics

Genomicsisthemostdifficult

ques8onhumanityhasevertakenon.

Beingpartofitisamazing.

Youarepartofthe0.0001%ofpopula8on

Enjoytheride!