User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/03/11

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Alien Genes New Primer Screen & TopTaq VS Takara R Taq

Reaction mixture conc. 1x 6 ' Reaction mixture conc. 1x 6
Template CCA3 1ng/ul 1 6 Template CCA3 1ng/ul 1 6
TopTaq buffer 10x 1 Takara R buffer 10x 1 6
dNTP mix 10 mM each 0.2 1.2 MgCl2 25mM 0.6 3.6
Primer mix 5 uM each 0.5 dNTP mix 2.5 mM each 0.8 4.8
TopTaq (QIAGEN) 5U/ul 0.05 0.3 Primer mix 5 uM each 0.5
Milli Q water 7.25 43.5 Takara R Taq 5U/ul 0.05 0.3
10 ul Milli Q water 6.05 36.3
PCR cycle 10 ul
94C 3 min PCR cycle
98C 10 sec 94C 30 sec
60C 3 min 60C 30 sec
x35 72C 1 min
72C 5 min x30

Load 5.0ul sample File:11march10.tif

AMP Lane Taq Mix Contents Fosmid
1 Top Taq AV10030 AV240_B
2 Top Taq AV10036 AV240_B
3 Top Taq AV10037 AV240_B
4 Top Taq AV10039 AV240_B
5 Top Taq POS Control AV10008
6 Top Taq no primer
7 Takara R AV10030 AV240_B
8 Takara R AV10036 AV240_B
9 Takara R AV10037 AV240_B
10 Takara R AV10039 AV240_B
11 Takara R POS Control AV10008
12 Takara R no primer