P20 | 0.5-20ul | 20ul | 0.5ul B.HowtoReadtheVolumeontheAutopipet Lookatthefrontfaceofthepipetandyouwillseeawindowwith3(three)digitsinside.ThediagrambelowshowstheMAXIMUMvaluethatcanorshouldbedialedinoneachsizepipet.Toexceedthesevalueswillputthepipetoutofcalibration.Besideeach\"window\"belowisthenumbersplaceitrepresents.Pleasetakethetimetolearnhowtoreadthemsoastoavoiddamagingthembydialingvaluesoutoftheirrange. GilsonP1000scalewindow. | P1000 | | P200,P100 | | P20 | | 1 | 1000\"s 100\"s 10\"s | 2,1 | 100\"s 10\"s 1\"s | 2 | 10\"s 1\"s 1decimal | 0 | 0 | 0 | 0 | 0 | 0 C.Howtochangetheloadvolume: Holdthepipetorhorizontallywiththeplungertoyourleft.Changeorsettheloadvolumebyturningtheknurledplasticknobatthetopofthehandgrip.Rollittowardsyoutodecreasethevaluesandawayfromyoutoincrease.WATCHCAREFULLYasyouchangethevaluestomakesureyoudon’ttakethenumbersoutofrange.Ifindoubt,reviewtheallowablevolumesforeachsizepipetor,andthenaskforhelp. D.Technique - GRIP:Holdthepipetorlikeaknifeinahorrorflickwithyourindexfingerontheplunger.Yourindexfingergivesyoumuchfinercontrolovertheplungeraction.
- LOADING:Loadasteriletip(blueforP1000;yellowforP200andallsmallersizesincludingVWRautopipets)andthenreclosethetipboxtomaintainsterilitiy.
- Pushtheplungerdownslowlytothepointoffirstresistance:thissetstheloadvolume.
- Whileholdingtheplungerattheloadvolumesetpoint,putthetipintothesolutionsothatitisimmersedjustenoughtocovertheend(3-4mm).
- Slowlyreleasetheplungertodrawuptheliquidmakingsuretokeepthetipimmersed.Visuallyinspecttheloadtomakesureitiscorrect.
- DELIVERY:Todeliverthevolume,placethetipintothereceivingvesselandpresstheplungerallthewaytothebottom-thisexpellsalltheliquidandgivesalittleextravolumetogetthelastdropletout-ANDTHEN,WITHOUTRELEASINGTHEPLUNGER,withdrawthetip.
- CHANGETIP?RepeatasnecessaryusingthesametipifNOTchangingsolutions.Useafreshtipforeverychangeofsolution,orwheneveritisprudenttomaintainsterileconditionsandavoidcrosscontaminationofsolutionsorcultures.
- TIPDISCHARGE:Whileholdingthetipoveranappropriatewastereceptacle,pressthewhitetipdischargeslideronthebackofthegrip.
| E.SmallVolumesTechnique:Withsmallvolumes,especiallythe1-10ulrangeusedinmolecularBIOLOGyprotocols,youmustkeeptrackofthedropletsyoupipet.Carefullyexpelltheliquiddropletonthesidewallofthetubesothatyoucanseeit. | |
Ifaddingtoalargervolume,flushthetipwiththesolventliquidafterexpellingthedroplettomakesureyougetallthedeliveryliquid.Withsmallvolumesyou\"llusuallyneedtocentrifugeandthenvortexthetubetogetagoodmixingofthereagents. F.ASimpleCheckforProperCalibration Checkthecalibrationofyourmicropipetbyusingthefactthat1ulofdeionized(ordistilled)waterhasamassof1ug.Pipetarangeofvolumesspanningthemicropipet\"susuablerangeandmassthemonatoploadingbalancehavingatleast3decimalplaceaccuracy.Pipetshavinggreaterthan0.1%errorshouldberecalibrated. G.InstructionsforUsingtheVWR5-50ulAutoPipets TheVWR5-50ulautopipetsfunctionsimilarlytotheGilsonPipetmanautopipets.Themajordifferenceliesinhowtosettheloadvolume.Thisisfurthercomplicatedbythelossofthe1ulindexring(whichsurroundstheplunger)fromseveralofthepipets. Theplungerhastwostops,thefirstbeingtheloadvolume,thesecondfordischargevolume.Thesepipetorsusethesamesmallyellowtips(yellowororangeboxes)asusedforP200andsmallerGilsonautopipets.Thetipejectorisadarkgraybuttonnearthetopofthehandgrip(seediagrambelow). Howtosettheloadvolume: Theplungerhasaverticallinedemarcatedbyhorizontalticksateach5ulincrementfrom0ulto50ul(toptobottom–seediagramatright).Whentheplungerisrotatedtoaligntheverticalmarkwiththe\"zero\"referencemarkontheindexring,theloadvolumeisthatofthehorizontallinejustshowingabovetheindexring.Inthediagramtotherightthatvolumeis10ul. Incrementsof1ulareachievedbyturningtheplungerCWtowardthe\"+\"symboltoincreasevolumeorCCWtowardthe\"-\"symboltodecreasevolume.Eachclickalignstheverticallinewithanewreferencemarkontheindexring.Toset13ul,goto10ulandthen3clicksCW. | Above:SideviewofVWR5-50ulautopipetsetto10ul. | Above:Pipetsetto20ul. | Onsomeofthepipetorstheindexringismissing.Tomaintaintheirutility,wehavemarkedthe1ulincrementsandthezeroreferencepointwithafelttiponthetopofthehandgrip.Thediagrambelowshowsthepipetorproperlysetfor10ul.Theindexringisequivalentindepthtothe5ulintervalsontheplunger.Theabsenceoftheindexringthereforeletsthe15ulmarkshowjustatthetopofthehandgrip.Toreadtheactualvolumeyou’llhavetosubtract5ulfromtheapparentvolume(15ulintheexample)indicatedatthetopofthehandgrip(seediagram).Askforhelpifyou’rehavingdifficultyreadingorsettingthecorrectloadvolume.
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