User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/06/18

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Nuclear Region 454 Amplification

Oleg Samples Amplified with primer no adapters

Extend with primers w/adapters


OlegNewExtraction.jpg


EX Takara RXN Mixture conc 1X 8 '
Milli Q water 6.15 49.2
Takara EX Buffer 10X 1 8
dNTP 2.5uM 0.8 6.4
Primer Mix 20uM 1
EX Taq 5U 0.05 0.4
Template Oleg PCR 1:10 1
64
8.0ul / RXN add 1.0ul Primer and 1.0ul PCR template
PCR cycle
98C 10 sec
60C 1 min
x5
AMP Lane Template Primer
1 Culture 11 B11
2 Culture 11 D11
3 Culture 11 E11
4 Culture 11 F11
5 Culture H B12
6 Culture H C12
7 Culture H E12
8 Culture H F12


Load 2.0ul of sample

18June10.jpg


Placed same PCR on machine for an additional 15 cycles


Load 2.0ul of sample

18June10B.jpg