2 × PCR Master mix (idafite irangi)
PCR Master mix ni ubwoko bwibisanzwe PCR yabanje gukemura yiteguye gukoresha, harimo Taq DNA Polymerase, DNTP ivanga MgCl2 na buffer nziza.Mugihe cyo kubyitwaramo, gusa primer nicyitegererezo birashobora kongerwaho kugirango byongerwe imbaraga, byoroshya cyane intambwe yo gukora igerageza.Ibicuruzwa birimo stabilisateur nziza kandi birashobora kubikwa amezi 3 kuri 4 ℃.Ibicuruzwa bya PCR bifite 3′-dA byerekana kandi birashobora gukoronizwa byoroshye muri T vector.
Ububiko
Ibicuruzwa bigomba kubikwa kuri -25 ℃ ~ -15 ℃ kumyaka ibiri.
Ibisobanuro
| Ubudahemuka (vs.Taq) | 1 × |
| Intangiriro ishyushye | No |
| Kurenga | 3′-A |
| Polymerase | Taq ADN Polymerase |
| Imiterere | SuperMix cyangwa Master mix |
| Umuvuduko wo Kwitabira | Bisanzwe |
| Ubwoko bwibicuruzwa | PCR Master mix (2 ×) |
Amabwiriza
1.Sisitemu yo Kwitabira
| Ibigize | Ingano (μL) |
| Icyitegererezo ADN | Birakwiriye |
| Primer 1 (10 μ mol / L) | 2 |
| Primer 2 (10 μ mol / L) | 2 |
| PCR Master mix | 25 |
| ddH2O | Kuri 50 |
2.Amasezerano yo kongera imbaraga
| Intambwe zizunguruka | Ubushyuhe (° C) | Igihe | Amagare |
| Mbere yo gutandukana | 94 ℃ | 5min | 1 |
| Gutandukana | 94 ℃ | 30 amasegonda | 35 |
| Annealing | 50-60 ℃ | 30 amasegonda | |
| Kwagura | 72 ℃ | 30-60sec / kb | |
| Kwagura kwanyuma | 72 ℃ | 10min | 1 |
Inyandiko:
1) Ikoreshwa ry'icyitegererezo: 50-200 ng ADN genomic;0.1- 10 ng ADN ya plasmid.
2) Mg2+kwibanda: Iki gicuruzwa kirimo mM 3 ya MgCl2 ikwiranye na PCR nyinshi.
3) Ubushyuhe bwa Annealing: Nyamuneka reba kuri Tm theoretical Tm agaciro ka Primers.Ubushuhe bwa annealing burashobora gushirwa kuri 2-5 ℃ munsi kurenza agaciro kamahame ya primer.
4) Igihe cyo kwagura: Kubiranga molekuline, 30 sec / kb birasabwa.Gukwirakwiza gene, 60sec / kb birasabwa.
Inyandiko
1.Kubwumutekano wawe nubuzima bwawe, nyamuneka wambare amakoti ya laboratoire hamwe na gants imwe ikoreshwa.
2.Gukoresha ubushakashatsi gusa!
