User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/15

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Contents

Nuclear Gene

Test amplification testing R Taq vs Top Taq

TOP TAQ RXN mixture conc. 1x 4 '
Template AV20ng/ul14
TopTaq buffer10x14
dNTP mix10 mM each0.20.8
Primer mix5 uM each1
TopTaq (QIAGEN)5U/ul0.050.2
Milli Q water6.7527
10 ul369.0ul / RXN
R Takara RXN Mixtureconc1X4
Milli Q water5.5522.2
Takara R Buffer10X14
MgCl225uM0.62.4
dNTP2.5uM0.83.2
Primer Mix5uM1
R Taq5U0.050.2
Template1ng / 20ng14
369.0ul / RXN
A PCR cycle
95C3 min
95C5 sec
60C1 min
x35
AMP Lane PCR Mix Template Primer
1R TaqAV 1ngAV10027
2R TaqAV 1ngA1
3R TaqAV 1ngA2
4R TaqAV 20ngAV10027
5R TaqAV 20ngA1
6R TaqAV 20ngA2
7Top TaqAV 1ngAV10027
8Top TaqAV 1ngA1
* AV10027 Positive Control


Image:15April10 B.tif

CCA3 Qiagen Extraction

~100-200 individuals

Followed standard Qiagen tissue extraction protocol

Nanodrop 0.5ng/ul DNA

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