2× PCR Premix （ine Dhayi） HYA442
2× PCR Master Mix ine Taq DNA Polymerase, dNTPs, nezvimwe zvinhu zvinodiwa nePCR. Master Mix inogara kwenguva refu kwemwedzi mitatu pa4℃ nema stabilizer edu akagadzirwa. Mushonga we pre-mix wakagadzirirwa PCR yakajairika uye wakagadzirira kushandiswa nekuwedzera DNA template nema primer. Zvigadzirwa zvePCR zvinogona kuiswa zvakananga kuti zvishandiswe ne electrophoresis ne bromophenol blue dye yakatoiswa kare. Zvigadzirwa zvakasimudzirwa zvine 3 '-dA protrusion uye zvinogona kuunganidzwa nyore nyore muT vector. 2× PCR Master Mix inoita kuti PCR iite nyore uye inoderedza kusvibiswa.
Zvikamu
| Zvikamu | Saizi-1 | Saizi-2 | Saizi-3 |
| 2 × PCR Premix (ine Dhayi) | 5mL | 1 mL × 15 | 1 mL × 50 |
Kuchengetera
Chengeta pa -20°C kwemakore maviri.
Mirayiridzo
1. Sisitimu yeKupindura
| Zvikamu | Vhoriyamu |
| 2 × PCR Premix (ine Dhayi) | 25 uL |
| Chinotangira Pamberi (10 μM) | 2 µL |
| Primer yekudzosera kumashure (10 μM) | 2 µL |
| Template yeDNA* | Chinoshanduka-shanduka |
| ddH2ZVINO | Kusvika ku50 µL |
*Kushandiswa kwetemplate: 50-200 ng genomic DNA; 0.1-10 ng plasmid DNA
2. Chirongwa Chekuita Nemhinduro
| Nhanho | Tembiricha | Nguva | Kutenderera |
| Kuchinja Kwekutanga | 94 ℃ | Maminitsi mashanu | 1 |
| Kuchinja kweganda | 94 ℃ | masekondi makumi matatu | 35 |
| Kunyungudutsa | 50-60 ℃ | masekondi makumi matatu | |
| Kuwedzera | 72 ℃ | 30-60 sec/kb | |
| Kuwedzerwa Kwekupedzisira | 72 ℃ | Maminitsi gumi | 1 |
| Bata | 4 ℃ | - | 1 |
1. Mg2+ huwandu: Chigadzirwa ichi chine 3 mM yeMgCl2, yakakodzera maitiro mazhinji ePCR.
2. Kupisa kwemvura: Ndapota tarisai kukosha kweTm kwema primers edzidziso. Kupisa kwemvura kunogona kuderedzwa kusvika pa2-5℃ pane kukosha kwemvura yedzidziso.
3. Nguva yekuwedzera: Kuti pave nekuziva mamorekuru, 30 sec/kb inokurudzirwa. Pakugadzira majini, 60 sec/kb inokurudzirwa.
Zvinyorwa
1. Zvigadzirwa zvePCR zvine 2× PCR Master Mix hazvina kukodzera polyacrylamide gel electrophoresis.
2. Kuti muve vakachengeteka uye vane hutano, ndapota pfekai majasi ekurabhoritari uye magirovhosi anoraswa kana muchivhiyiwa.
3. Inoshandiswa pakutsvaga chete!