Ungayisebenzisa kanjani i-pipte omncane amavolumu ane-paipette yezandla eziphathwayo

Njengoba kugxilwe kakhulu ekunciphiseni ama-reagents nezindleko, amavolumu amancane afunwa kakhulu, isib, ukulungiswa kwe-PCR Mastermix noma ukuphendula kwe-enzyme. Kepha i-Pipetting amanani amancane asuka ku-0,2 - 5 μL abeka izinselelo ezintsha zokunemba nokunemba kwePipetting. Amaphuzu alandelayo abalulekile:

1. I-PIPETE NE-PIPE SIPE: Njalo khetha i-Pipette ngevolumu eliphansi kakhulu elisezingeni eliphakeme kanye nethiphu elincane kunazo zonke ukugcina i-cushion yomoya incane ngangokunokwenzeka. Lapho i-Pipetting 1 μl isib
2. Ukulinganisa nokulungiswa: Kubalulekile ukuthi ama-Pipete akho alinganiselwe futhi agcinwe. Ukulungiswa okuncane nezingxenye eziphukile kwi-pipette kuholele ekunyuseni okukhulu kumanani wephutha elihleliwe. Ukulinganisa ngokuya nge-ISO 8655 kumele kwenziwe kanye ngonyaka.
3. Ama-Pipette asuswayo afanele: Bheka ukuthi ngabe une-pipette enhle yokufuduka ngenani eliphansi levolumu kwilebhu yakho. Ngokuvamile, ukusebenzisa lolu hlobo lwe-pipette kuholela emthethweni ongcono wepayiphu ngokuya ngokunemba nokunemba kunokuthola amapayipi e-classic air-cushion.
4. Zama ukusebenzisa amavolumu amakhulu: ungacabanga ukunciphisa isampula yakho kumanani amakhulu wePipette ngenani elifanayo ekuphenduleni kokugcina. Lokhu kunganciphisa amaphutha ePipetting nge-fafaza kakhulu yesampula.

Ngaphezu kwethuluzi elihle, umcwaningi kufanele abe necebo elihle kakhulu lePipetting. Naka ngokukhethekile izinyathelo ezilandelayo:

1. I-Tip Okunamathiselwe: Musa ukujaha i-pipette esiphuzeni njengoba lokhu kungalimaza ukuphela kwethiphu enhle ngoba ugongolo oluwuketshezi ukuze luqondiswe noma lulimaze yi-orifice. Faka isicelo sengcindezi yokukhanya kuphela lapho unamathisela ithiphu bese usebenzisa i-pipette ngesigaxa esilayishwe entwasahlobo.
2. Ukubamba i-PIPETTE: Musa ukubamba i-pipette esandleni sakho ngenkathi ulinde i-centrifuge, i-cypeer, njll. Ingaphakathi le-pipette lizofudumeza futhi lihole i-air cushion ukukhulisa okuholela ekuphambukeni kwevolumu esethiwe lapho ipipeting.
3. I-Pre-Whetting: Umswakama womoya ngaphakathi kwethiphu ne-pipette ulungiselela ithiphu lesampula futhi uvikele ukuhwamuka lapho efisa ivolumu yokudlulisa.
4. Isifiso esiqondile: Lokhu kubaluleke kakhulu lapho uphatha amavolumu amancane ukugwema umphumela we-capillary owenzeka lapho i-pipette ibanjelwa engela.
5. Ukujuliswa kokucwiliswa: Gxila ithiphu kancane ngangokunokwenzeka ukuvikela uketshezi olungena kwi-tip ngenxa yomphumela we-capillary. Umthetho wesithupha: incane ithiphu nevolumu, kwehlisa ukujula kokucwiliswa. Sincoma ubukhulu be-2 mm lapho kuthola amanani amancane.
6. Ukusabalalisa ngo-45 ° Angle: Ukugeleza okuphelele koketshezi kuqinisekisiwe lapho i-pipette ibanjelwe e-45 ° angle.
7. Oxhumana naye odongeni lwe-Vessel noma indawo ewuketshezi: Amavolumu amancane angasatshalaliswa kahle lapho ithiphu libanjwe odongeni lomkhumbi, noma olufakwe luketshezi. Ngisho nokwehla kokugcina okuvela esiqongweni kungahanjiswa ngokunembe.
8. Blow-Out: Ukuqhuma kuyaphoqeleka ngemuva kokuhambisa amavolumu aphansi ukuze kukhiphe ngisho nethonsi lokugcina le-ketshezi elikhona empini. Ukuqhuma kufanele futhi kwenziwe ngokulwa nodonga lomkhumbi. Qikelela ukuthi ungalethi ama-bubble omoya kusampula lapho wenza ukushaya okuphuma endaweni ewuketshezi.