2 × PCR Super Mix (ine Dhayi)
2 × PCR Master Mix ine Taq DNA Polymerase, dNTPs, uye zvimwe PCR-inodiwa zvikamu.Iyo Master Mix yakagadzikana kwemwedzi mitatu pa4 ℃ ine edu akagadziridzwa stabilizers.Iyo pre-musanganiswa mhinduro yakagadziridzwa kune yakajairwa PCR uye yakagadzirira kushandisa nekuwedzera DNA template uye primers.Zvigadzirwa zvePCR zvinogona kutakurwa zvakanangana ne electrophoresis ine pre-yakatakura bromophenol dhayi yebhuruu.Zvigadzirwa zvakakwidziridzwa zvine 3 '-dA protrusion uye zvinogona kugadzirwa nyore muT vector.Iyo 2 × PCR Master Mix inorerutsa maitiro ePCR uye inoderedza kusvibiswa.
Storage Conditions
Zvigadzirwa zvinofanirwa kuchengetwa pa -25 ℃ ~ -15 ℃ kwemakore maviri.
Zvinotsanangurwa
| Kuvimbika (vs.Taq) | 1× |
| Sravana Sameeralu Serial 4th Hot Start | No |
| Overhang | 3 '-A |
| Polymerase | Taq DNA Polymerase |
| Reaction Format | SuperMix kana Master Mix |
| Reaction Speed | Standard |
| Product Type | PCR Master Mix (2x) |
Mirayiridzo
1.Reaction System
| Zvikamu | Volume(μL) |
| DNA template | Inokodzera |
| Primer 1 (10 μmol/L) | 2 |
| Primer 2 (10 μmol/L) | 2 |
| 2 × PCR Master Mix | 25 |
| ddH2O | kusvika ku50 |
2.Amplification Protocol
| Matanho ekutenderera | Tembiricha (°C) | Nguva | Cycles |
| Initial denaturation | 94 | 5 min | 1 |
| Denaturation | 94 | 30 sec | 35 |
| Annealing | 50-60 | 30 sec | |
| Extension | 72 | 30-60 sec/kb | |
| Final extension | 72 | 10 min | 1 |
Cherechedza:
1) Template kushandiswa: 50-200ng genomic DNA;0.1-10ng plasmid DNA.
2) Mg2+concentration: Ichi chigadzirwa chine 3mM yeMgCl2, yakakodzera kune akawanda PCR maitiro.
3) Anealing tembiricha: Ndokumbira utarise kune theoretical Tm kukosha kwekutanga.Iyo annealing tembiricha inogona kusetwa ku2-5 ℃ yakaderera pane theoretical kukosha kweprimer.
4) Nguva yekuwedzera: Nekuzivikanwa kwema molecular, 30 sec / kb inokurudzirwa.Kune gene cloning, 60 sec/kb inokurudzirwa.
Notes
1.Zvigadzirwa zvePCR zvine 2 × PCR Master Mix hazvina kukodzera polyacrylamide gel electrophoresis.
2.Nekuchengetedza uye hutano hwako, ndapota pfeka majasi erabhu uye magurovhosi anoraswa kuti ushande.
3.Inoshandiswa pakutsvagisa CHETE!
