Ngwa a na-eji sistemu nchekwa lysis pụrụ iche nke nwere ike hapụ RNA ngwa ngwa site na nlele cell emebere maka mmeghachi omume RT-qPCR, si otú a na-ewepụ usoro ịdị ọcha nke RNA na-ewe ma na-arụsi ọrụ ike.Enwere ike nweta ndebiri RNA n'ime naanị nkeji 7.Ngwakọta 5 × Direct RT Mix na 2 × Direct qPCR Mix-SYBR reagents nke ngwa ahụ na-enye nwere ike nweta nsonaazụ PCR ozugbo na nke ọma.

5 × Direct RT Mix na 2 × Direct qPCR Mix-SYBR nwere nnabata inhibitor siri ike, enwere ike iji lysate nke ihe nlele ahụ dị ka ndebiri maka RT-qPCR ozugbo.Ngwa a nwere RNA pụrụ iche Foregene reverse transcriptase, yana Hot D-Taq DNA polymerase, dNTPs, MgCl.₂, ihe nchekwa mmeghachi omume, PCR optimizer na stabilizer.

200 × 20μl Rxns, 1000 × 20μl Rxns

∎ Dị mfe ma dị irè: site na teknụzụ Cell Direct RT, enwere ike nweta ihe nlele RNA n'ime naanị nkeji asaa.

■ Ọchịchọ nlele dị ntakịrị, ebe ọ bụ na enwere ike ịnwale mkpụrụ ndụ 10.

∎ Mgbakwunye dị elu: ọ nwere ike ịchọpụta RNA ngwa ngwa n'ime sel ndị emebere na 384, 96, 24, 12, 6-well plates.

∎ DNA Eraser nwere ike wepụ ngwa ngwa genomes ewepụtara, belata mmetụta dị na nsonaazụ nnwale na-esote.

∎ Sistemu RT na qPCR emezigharịrị na-eme ka ntụgharị ntụgharị RT-PCR nzọụkwụ abụọ rụọ ọrụ nke ọma yana PCR akọwapụtara nke ọma, ma na-eguzogide ndị na-egbochi mmeghachi omume RT-qPCR.

Oke ngwa: mkpụrụ ndụ omenala.

- RNA wepụtara site na nlele lysis: naanị ọdabara na ndebiri RT-qPCR nke ngwa a.

- Enwere ike iji ngwa ahụ maka ebumnuche ndị a: nyocha okwu mkpụrụ ndụ ihe nketa, nkwenye nke mmetụta ịgbachitere mkpụrụ ndụ ihe nke siRNA, nyocha ọgwụ, wdg.