2 × PCR Master Mix (isina Dhayi)
PCR Master Mix imhando yeyakajairwa PCR premixed mhinduro yakagadzirira kushandisa, kusanganisira Taq DNA Polymerase, dNTP musanganiswa MgCl2 uye yakagadziridzwa buffer.Munguva yekuita, chete primer uye template inogona kuwedzerwa kuti iwedzere, iyo inorerutsa zvakanyanya matanho ekushanda ekuedza.Ichi chigadzirwa chine yakanakisa stabilizers uye chinogona kuchengetwa kwemwedzi mitatu pa4 ℃.Chigadzirwa chePCR chine 3'-dA protrusion uye inogona kugadzirwa nyore muT vector.
Storage Conditions
Chigadzirwa chacho chinofanira kuchengetwa pa -25 ℃ ~ -15 ℃ kwemakore maviri.
Zvinotsanangurwa
| Kutendeseka (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 (2×) |
Mirayiridzo
1.Reaction System
| Zvikamu | Saizi (μL) |
| DNA template | Inokodzera |
| Primer 1 (10 μmol/L) | 2 |
| Primer 2 (10 μmol/L) | 2 |
| PCR Master Mix | 25 |
| ddH2O | kusvika ku50 |
2.Amplification Protocol
| Matanho ekutenderera | Tembiricha (°C) | Nguva | Cycles |
| Pre-denaturation | 94 ℃ | 5 mins | 1 |
| Denaturation | 94 ℃ | 30 sec | 35 |
| Annealing | 50-60 ℃ | 30 sec | |
| Extension | 72 ℃ | 30-60sec/kb | |
| Final Extension | 72 ℃ | 10mins | 1 |
Notes:
1) Template kushandiswa: 50-200 ng genomic DNA;0.1- 10 ng plasmid DNA.
2) Mg2+concentration: Ichi chigadzirwa chine 3 mM yeMgCl2 yakakodzera kune akawanda PCR maitiro.
3) Anealing tembiricha: Ndokumbira utarise kune theoretical Tm kukosha kwePrimers.Iyo annealing tembiricha inogona kusetwa kusvika 2-5 ℃ yakaderera pane theoretical kukosha kweprimer.
4) Nguva yekuwedzera: Nekuzivikanwa kwema molecular, 30 sec / kb inokurudzirwa.Nezve gene cloning, 60sec/kb inokurudzirwa.
Notes
1.Nekuchengetedza uye hutano hwako, ndapota pfeka majasi erabhu uye magurovhosi anoraswa kuti ushande.
2.Yekushandisa tsvakurudzo chete!
