TAIL PCR反应体系和程序
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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
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第一次扩增
1st amplification
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第二次扩增
2ed amplification
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第三次扩增
3rd amplification
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92 ℃3min
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94 ℃30min ＊＊＊
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94 ℃10s
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95 ℃1min
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58 ℃1min ＊＊＊
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94 ℃30s ＊＊＊＊
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94 ℃30 s ＊
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72 ℃2min ＊＊＊
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42 ℃30s ＊＊＊＊
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62 ℃1min ＊
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94 ℃30s ＊＊＊
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72 ℃2min ＊＊＊＊
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72 ℃2min ＊
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58 ℃1min ＊＊＊
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72 ℃5min
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94 ℃30s
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72 ℃2min ＊＊＊
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25 ℃3min
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94 ℃30s ＊＊＊
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Ramp to72℃over 3min
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44 ℃1min ＊＊＊
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72 ℃2min
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72 ℃2min ＊＊＊
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|
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94 ℃30s ＊＊
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72 ℃5min
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|
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44 ℃1min ＊＊
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|
|
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72 ℃2min ＊＊
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94 ℃30s ＊＊＊
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62 ℃1min ＊＊＊
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72 ℃2min ＊＊＊
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94 ℃30s ＊＊＊
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62 ℃2min ＊＊＊
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72 ℃2min ＊＊＊
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|
|
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94 ℃30s ＊＊＊
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|
|
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44 ℃1min ＊＊＊
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|
|
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72 ℃2min ＊＊＊
72 ℃5min
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注： ＊，＊＊，＊＊＊，＊＊＊＊分别代表x5 ，x10 ，x15 ，x30 循环
