User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/06/08

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Contents

Nuclear Gene Adenita Cultures 454 Amplification

A. ricciae

EX Takara RXN Mixture conc 1X 5
Milli Q water4.1520.75
Takara EX Buffer10X15
dNTP2.5uM0.84
Primer Mix20uM1
EX Taq5U0.050.25
TemplateInstagene315
45
9.0ul / RXN add 1.0ul Primer
PCR cycle
98C10 sec
60C1 min
x35
AMP LaneTemplatePrimer
1A. RicciaeB3
2A. RicciaeC3
3A. RicciaeD3
4A. RicciaeE3
5A. RicciaeF3
6Neg ControlB3
7Neg ControlC3
8Neg ControlD3
9Neg ControlE3
10Neg ControlF3
11Pos ControlB3
12Pos ControlC3
13Pos ControlD3
14Pos ControlE3
15Pos ControlF3
*Positive Control = AV 20ng

Loaded 2.0ul Image:08June10.jpg

HCO/LCO Adenita Culture Cycle Sequencing

For failed sequencing sample 14

CS Reaction mixture conc. 1x ' ' AMP Lane Template ' Sample Number Primer
Big Dye Terminator0.51Ithaca 1A314HCO
Primer (FWD or RVS)5uM0.42Ithaca 1A314LCO
DMSO0.1
5X Seq Buffer5X0.4
Water0.6
Template *~5ng/ul4
2.0ul / RXN
*QIAquick Cleaned PCR product
Cycle Seq Program
96C10 sec
50C5 sec
60C4 min
x60
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