Pytorch中的RNN之pack_padded_sequence()和pad_packed ...
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SonicationofBacteria
Thisprocedureisusedtobreakcellsthatcontainaproteintobepurified.ThisprocedureisvalidonlyfortheuseonAppling"slabsonicatior.
PREPARATION
- Placeyoursampleina15mlelongatedplastictubesuchthatthesample-tubewallinterfaceareaismaximized.
- Fillasmallcontainerwithiceandsalt(NaCl)andthenaddasmallamountofwater(thesaltlowersthefreezingtemp.).
- Placethetubewiththesampleintheicecontainermakingsurethattheicebathcompletlycoversthewallsofthetube.
SONICATION
- Turnonthesonicatorandcalibratebyplacingthe20/40switchinthe20possitionandthecontrolpowerswitchinthe60postion.Starttheapparatusbypressingthefootpedalandmovethecalibrationknobsuchthattheoutputsignalisataminimun.
- Movethepowerswitchtopthe100positionandrepeatabovecalibrationprocedure.
- Cleantheprobewithethanol,letdry,andplaceitinsidethetube.Makesurethatthecoldtubeisinsidetheicecontainer.Movethe20/40switchtothe40positionandthepowerswitchto70.Wearyourprotectiveheadfonesandstartthesonication.Leavethesonicatoronfor30secondsandthenreleasethefootpedal.
- Waitaboutoneortwominutesuntilthetubecoolsdownagainandgentlyshakethetube.Placeitoniceandrepeattheexperiment.
- Movethepowerswitchto90andrepeattheexperiment.Thesequenceofsonicationis2Xat70for30secondsand1Xat90for30secondswaiting1-2minutesontheicebathinbetween.
Note:Someproteinsresistsonicationmorethanotherssodifferenttimesmaybeneeded.Theinitialcellsampleisviscousandasthesonicationprocesstheviscosityofthesampledecreasesduetomacromolecularshearing.AnotherwaytomonitortheamountofsonicationneededtobreakopenthebacteriaistomonitorthereleaseofDNAandRNAbymeasuringtheabsorbanceat260nmofyoursampleaftereverysonicationstep.TheA260willincreaseuntilitreachesaplateau--thismeansallthebacteriaarelysedandnomorenucleicacidscanbeliberated.Alwaysremembertocooldownyoursampleinbetweenandaftersoniicationtopreventheatdenaturingyourprotiens
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