User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/06

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Contents


Nuclear Gene HS Takara PCR Optimization

HS Takara RXN Mixture 3.0ul Instagene conc 1X 5.5 '
Milli Q water3.117.05
Takara HS Buffer10X211
dNTP2.5uM0.84.4
Primer Mix20uM1
HS Taq5U0.10.55
TemplateInstagene316.5
49.59.0ul / RXN
add 1.0ul Primer
HS Takara RXN Mixture 4.0ul Instageneconc1X5.5
Milli Q water2.111.55
Takara HS Buffer10X211
dNTP2.5uM0.84.4
Primer Mix20uM4
HS Taq5U0.10.55
TemplateInstagene422
49.59.0ul / RXN
add 1.0ul Primer
A PCR cycle
98C10 sec
60C1 min
x35
AMP LaneTemplatePrimerAmt of Template
13 InstageneA23
23 InstageneA33
33 InstageneA43
43 InstageneA53
53 InstageneA63
64 InstageneA23
74 InstageneA33
84 InstageneA43
94 InstageneA53
104 InstageneA63
113 InstageneA24
123 InstageneA34
133 InstageneA44
143 InstageneA54
153 InstageneA64
164 InstageneA24
174 InstageneA34
184 InstageneA44
194 InstageneA54
204 InstageneA64
21AV 2ngA21
22AV 2ngA31
23AV 2ngA41
24AV 2ngA51
25AV 2ngA61
*AV 1ng Positive Control

Load 2.0ul Sample

Image:06May10.jpg


AV 1ng control failed - bad DNA dilution

Dye Dilution Test

1:10 Dye Stock : MilliQ Water

From 06May10 PCR - samples 1-10 Image:dye test.jpg

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