TAIL PCR反应体系和程序
TAIL PCR反应体系和程序TAIL-PCR 反应体系(20μl体系)Templet 1μl10x buffter 2.0μlMg 2+ 1.5μldNTP
TAIL PCR反应体系和程序
TAIL-PCR 反应体系(20μl体系)
Templet 1μl
10x buffter 2.0μl
Mg 2+ 1.5μl
dNTP 0.2μl (10mmol)
P1 0.2μl (15μmol)
P2(AD) 1.3μl (30μmol)
Taq 0.2μl (5U/μl)
ddH 2 O 15.1μl
Tail-PCR 反应程序及条件
Program used for Tail-PCR amplification
第一次扩增 1st amplification | 第二次扩增 2ed amplification | 第三次扩增 3rd amplification |
92 ℃3min | 94 ℃30min ＊＊＊ | 94 ℃10s |
95 ℃1min | 58 ℃1min ＊＊＊ | 94 ℃30s ＊＊＊＊ |
94 ℃30 s ＊ | 72 ℃2min ＊＊＊ | 42 ℃30s ＊＊＊＊ |
62 ℃1min ＊ | 94 ℃30s ＊＊＊ | 72 ℃2min ＊＊＊＊ |
72 ℃2min ＊ | 58 ℃1min ＊＊＊ | 72 ℃5min |
94 ℃30s | 72 ℃2min ＊＊＊ | |
25 ℃3min | 94 ℃30s ＊＊＊ | |
Ramp to72℃over 3min | 44 ℃1min ＊＊＊ | |
72 ℃2min | 72 ℃2min ＊＊＊ | |
94 ℃30s ＊＊ | 72 ℃5min | |
44 ℃1min ＊＊ | ||
72 ℃2min ＊＊ | ||
94 ℃30s ＊＊＊ | ||
62 ℃1min ＊＊＊ | ||
72 ℃2min ＊＊＊ | ||
94 ℃30s ＊＊＊ | ||
62 ℃2min ＊＊＊ | ||
72 ℃2min ＊＊＊ | ||
94 ℃30s ＊＊＊ | ||
44 ℃1min ＊＊＊ | ||
72 ℃2min ＊＊＊ 72 ℃5min | ||
注： ＊，＊＊，＊＊＊，＊＊＊＊分别代表x5 ，x10 ，x15 ，x30 循环
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