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1、2006-4raimi编辑整理於DICPSDS-PAGE"HallofShame"Youmayverywellhavepreparedanearlyperfectgel,andwouldhaveadifficulttimeimprovingupontheproduct.Ifthatisso,thenbyallmeansgloataboutit!Ifyoudidn'tdosuchahotjobdon'tdespair.Allgoodscientistslearnfromtheirmistakes,infact,withou
2、tmakingmistakesmostofuswouldn'tlearnmuchatall!The"HallofShame"presentsexamplesofsomeoftheworstgelsstudents(andinstructor)haveruninpastlabs,withanexampleortwofromaresearchlab.Theyrepresentmanyofthewaysonecanmessupagel(butnotallofthem-we'restillfindingnewways!).See
3、whatfeaturesofyourowngel(s)wereunsatisfactory-oratleastlessthanperfect-andusetheillustrationstofigureoutwhatyoumightdotoimproveyourtechnique.Tocritiqueyourownworkidentifyyoursymptomsandusethegallerytoselectappropriateexample(s).Eachexampleislinkedtoafullsizedimag
4、ewithsuggesstedexplanationsforthesymptoms.Fromthis"hallofshame"youshouldbeabletodeterminewhatcanbedonetocorrecttheproblem(s).Ifyouwishyoumaybrowsebydescriptionsofsymptoms,listedbelowthegallery.Symptoms1—SmearsSmearedgels–example1Smearingcanhaveavarietyofcauses,bu
5、tmostcommonlyitisduetoanunevenlypouredacrylamidemixtureorduetogrossoverloadingofprotein.Inthisexamplethegelwasnotproperlypoured,sothatthelowerhalfhadbeguntopolymerizebeforetheupperpartwaspoured.Thefirstgelmixbegantopolymerizetooquickly.Ratherthanprepareanewgelcas
6、sette,thestudentssimplystoppedpouring,preparedanewmix,andpoureditontopoftheold.Obviously,thebondwasn'tparticularlygood.Smearedgels–example2Inthisexamplealotofproteinwasloadedineachofthewells,allthewayacross.Mostofthelanescanstillbeinterpreted,butsmearingisparticu
7、larlyevidentinlanes3and4.Thoselanescontainedprimarilyoneprotein(hemoglobinsubunit),asdidlanes9and10.However,thegroupthatloaded3and4underestimatedtheproteinconcentrationsintheirredcelllysateandredcellcytoplasmfractions.Thosefractionscontainedsomuchproteinthatsampl
8、eshadtobedilutedpriortopreparingaproteinassaytube.Thestudentsforgottotakethedilution1原始网址：http://www.ruf.rice.edu/~bioslabs/studies/sds-page/sdsgoofs.html2006-
