2×PCR Super Mix (pẹlu Dye)
2× PCR Master Mix ni Taq DNA Polymerase, dNTPs, ati awọn paati PCR miiran ti o nilo.Adalu Titunto jẹ iduroṣinṣin fun awọn oṣu 3 ni 4℃ pẹlu awọn amuduro adani wa.Ojutu iṣaju iṣaju jẹ iṣapeye fun PCR ti aṣa ati ṣetan lati lo nipa fifi awoṣe DNA kun ati awọn alakoko.Awọn ọja PCR le ṣe kojọpọ taara fun electrophoresis pẹlu awọ buluu bromophenol ti a ti ṣajọ tẹlẹ.Awọn ọja imudara ni 3 '-dA protrusion ati pe o le ni irọrun cloned sinu T fekito.Mix 2 × PCR Titunto simplifies ilana PCR ati dinku ibajẹ.
Awọn ipo ipamọ
Awọn ọja yẹ ki o wa ni ipamọ ni -25 ℃ ~ -15 ℃ fun 2 ọdun.
Awọn pato
| Iduroṣinṣin (vs.Taq) | 1× |
| Gbona Bẹrẹ | No |
| Overhang | 3'-A |
| Polymerase | Taq DNA Polymerase |
| Ifaseyin kika | SuperMix tabi Titunto Mix |
| Iyara lenu | Standard |
| Ọja Iru | PCR Titunto Mix (2x) |
Awọn ilana
1.ifaseyin System
| Awọn eroja | Iwọn didun(μL) |
| DNA awoṣe | Dara |
| Alakoko 1 (10 μmol/L) | 2 |
| Alakoko 2 (10 μmol/L) | 2 |
| 2× PCR Titunto Mix | 25 |
| ddH2O | si 50 |
2.Ilana Imudara
| Awọn igbesẹ yipo | Iwọn otutu (°C) | Aago | Awọn iyipo |
| Ibẹrẹ denaturation | 94 | 5 min | 1 |
| Denaturation | 94 | 30 iṣẹju-aaya | 35 |
| Annealing | 50-60 | 30 iṣẹju-aaya | |
| Itẹsiwaju | 72 | 30-60 iṣẹju-aaya / kb | |
| Ipari ipari | 72 | 10 min | 1 |
Akiyesi:
1) Lilo awoṣe: 50-200ng DNA genomic;0.1-10ng DNA pilasima.
2) mg2+ifọkansi: Ọja yii ni 3mM ti MgCl2, o dara fun ọpọlọpọ awọn aati PCR.
3) otutu Annealing: Jọwọ tọka si iye Tm imọ-jinlẹ ti awọn alakoko.Iwọn otutu mimu le ṣee ṣeto si 2-5℃ kekere ju iye imọ-jinlẹ ti alakoko lọ.
4) Akoko itẹsiwaju: Fun idanimọ molikula, 30 iṣẹju-aaya / kb ni a ṣe iṣeduro.Fun ẹda ẹda, 60 iṣẹju-aaya/kb ni iṣeduro.
Awọn akọsilẹ
1.Awọn ọja PCR pẹlu 2× PCR Master Mix ko dara fun polyacrylamide gel electrophoresis.
2.Fun ailewu ati ilera rẹ, jọwọ wọ awọn aṣọ laabu ati awọn ibọwọ isọnu fun iṣẹ.
3.O ti lo fun lilo iwadi NIKAN!
