2 × PCR Master Mix (ba tare da Dye ba)
PCR Master Mix wani nau'in bayani ne na PCR na al'ada wanda ke shirye don amfani, gami da Taq DNA Polymerase, dNTP mix MgCl2 da ingantaccen buffer.A lokacin amsawa, kawai za a iya ƙara firamare da samfuri don haɓakawa, wanda ke sauƙaƙe matakan aiki na gwaji sosai.Wannan samfurin yana ƙunshe da ingantattun masu daidaitawa kuma ana iya adana su na tsawon watanni 3 a 4 ℃.Samfurin PCR yana da fitowar 3′-dA kuma ana iya haɗa shi cikin sauƙi cikin T vector.
Yanayin Ajiya
Ya kamata a adana samfurin a -25 ℃ ~ -15 ℃ na shekaru biyu.
Ƙayyadaddun bayanai
| Aminci (vs.Taq) | 1× |
| Zafafan Farawa | No |
| Tsayawa | 3'- A |
| Polymerase | Taq DNA Polymerase |
| Tsarin martani | SuperMix ko Master Mix |
| Saurin amsawa | Daidaitawa |
| Nau'in Samfur | PCR Master Mix (2×) |
Umarni
1.Tsarin martani
| Abubuwan da aka gyara | Girman (μL) |
| Samfurin DNA | Dace |
| Farko 1 (10 μmol/L) | 2 |
| Farko 2 (10 μmol/L) | 2 |
| PCR Master Mix | 25 |
| ddH2O | Zuwa 50 |
2.Ƙarfafa Ƙarfafawa
| Matakan zagayowar | Zazzabi (°C) | Lokaci | Zagaye |
| Pre-denaturation | 94 ℃ | 5 min | 1 |
| Denaturation | 94 ℃ | 30 seconds | 35 |
| Annealing | 50-60 ℃ | 30 seconds | |
| Tsawaita | 72 ℃ | 30-60 seconds/kb | |
| Ƙarshe Ƙarshe | 72 ℃ | Minti 10 | 1 |
Bayanan kula:
1) Yin amfani da samfur: 50-200 ng DNA na genomic;0.1-10 ng DNA plasmid.
2) Mg2+maida hankali: Wannan samfurin ya ƙunshi 3 mM na MgCl2 dace da yawancin halayen PCR.
3) Zazzage zafin jiki: Da fatan za a koma zuwa ƙimar Tm na ka'idar.Za'a iya saita zafin jiki na annealing zuwa 2-5 ℃ ƙasa da ƙimar ƙa'idar farko.
4) Lokacin tsawaitawa: Don gano kwayoyin halitta, ana ba da shawarar 30 sec/kb.Don cloning gene, 60sec/kb ana bada shawarar.
Bayanan kula
1.Don amincin ku da lafiyar ku, da fatan za a sa riguna na lab da safofin hannu masu zubarwa don aiki.
2.Don amfani da bincike kawai!
