Setsi sena se sebelisa sistimi e ikhethileng ea lysis buffer e ka lokollang RNA kapele ho lisampole tsa lisele tse hlahisitsoeng bakeng sa karabelo ea RT-qPCR, ka hona e felisa ts'ebetso ea tlhoekiso ea RNA e jang nako le e boima.Setšoantšo sa RNA se ka fumanoa ka metsotso e 7 feela.The 5×Direct RT Mix le 2×Direct qPCR Mix-SYBR reagents e fanoeng ke kit e ka fumana kapele le ka nepo liphetho tsa nako ea nnete tsa PCR.

5 × Direct RT Mix le 2 × Direct qPCR Mix-SYBR li na le mamello e matla ea inhibitor, 'me lysate ea lisampole e ka sebelisoa e le template ea RT-qPCR ka ho toba.Setsi sena se na le RNA e phahameng ea kamano e phahameng ea Foregene reverse transcriptase, le Hot D-Taq DNA polymerase, dNTPs, MgCl.₂, buffer ea karabelo, PCR optimizer le stabilizer.

200×20μl Rxns, 1000×20μl Rxns

■ E bonolo ebile e sebetsa : ka theknoloji ea Cell Direct RT, lisampole tsa RNA li ka fumanoa ka metsotso e 7 feela.

■ Tlhokahalo ea sampole e nyane, e ka tlase ho lisele tse 10 li ka lekoa.

■ Phallo e phahameng: e khona ho lemoha ka potlako RNA liseleng tse entsoeng ka lipoleiti tse 384, 96, 24, 12, 6.

■ DNA Eraser e ka tlosa ka potlako liphatsa tsa lefutso tse lokollotsoeng, ea fokotsa haholo tšusumetso liphethong tse latelang tsa liteko.

■ Optimized RT le qPCR system e etsa hore RT-PCR reverse tranescript e sebetse hantle le hore PCR e be e hlakileng haholoanyane, 'me e hanane le RT-qPCR reaction inhibitors.

Sekhahla sa ts'ebeliso: lisele tse tsoetseng pele.

- RNA e lokollotsoe ka mohlala oa lysis: e sebetsa feela ho template ea RT-qPCR ea kit ena.

- Thepa e ka sebelisoa bakeng sa merero e latelang: tlhahlobo ea polelo ea liphatsa tsa lefutso, netefatso ea siRNA-mediated gene silence effect, tlhahlobo ea lithethefatsi, joalo-joalo.