Chidachi chimagwiritsa ntchito njira yapadera ya lysis buffer yomwe imatha kumasula mwachangu RNA kuchokera ku zitsanzo zama cell otukuka pamachitidwe a RT-qPCR, potero kuchotsa njira yoyeretsera ya RNA yomwe imatenga nthawi komanso yovuta.The RNA template ikhoza kupezeka mu mphindi 7 zokha.5 × Direct RT Mix ndi 2 × Direct qPCR Mix-SYBR reagents zoperekedwa ndi zida zitha kupeza mwachangu komanso moyenera zotsatira za PCR zenizeni zenizeni.

5 × Direct RT Mix ndi 2 × Direct qPCR Mix-SYBR ali ndi kulekerera kwamphamvu kwa inhibitor, ndipo ma lysate a zitsanzo angagwiritsidwe ntchito ngati template ya RT-qPCR mwachindunji.Chidachi chili ndi RNA yapamwamba kwambiri ya Foregene reverse transcriptase, ndi Hot D-Taq DNA polymerase, dNTPs, MgCl.₂, zotchingira zochita, PCR optimizer ndi stabilizer.

200×20μl Rxns, 1000×20μl Rxns

■ Zosavuta komanso zothandiza : ndiukadaulo wa Cell Direct RT, zitsanzo za RNA zitha kupezeka m'mphindi 7 zokha.

■ Zofuna zachitsanzo ndizochepa, mpaka ma cell 10 akhoza kuyesedwa.

■ Kutulutsa kwakukulu: kumatha kuzindikira mwachangu RNA m'maselo opangidwa mu 384, 96, 24, 12, 6-chitsime mbale.

■ DNA chofufutira akhoza kuchotsa mwamsanga matupi athu omasulidwa, kwambiri kuchepetsa zotsatira zoyeserera.

■ Makina okhathamiritsa a RT ndi qPCR amapangitsa kuti masitepe awiri a RT-PCR alembedwe bwino kwambiri komanso PCR yachindunji, komanso yosagwirizana ndi RT-qPCR reaction inhibitors.

Kuchuluka kwa ntchito: maselo otukuka.

- RNA yotulutsidwa ndi zitsanzo za lysis: imagwira ntchito pa template ya RT-qPCR ya zida izi.

- Zidazi zitha kugwiritsidwa ntchito pazifukwa izi: kusanthula kwa jini, kutsimikizira kwa siRNA-mediated gene silence effect, kuyang'anira mankhwala, etc.