User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/05/07

From OpenWetWare

Jump to: navigation, search
Alien genes Main project page
Previous entry      Next entry

Contents


Nuclear Region HS Takara PCR Optimization

Test 4.0ul of 5.0uM Primer Concentration

HS Takara RXN Mixture 3.0ul Instagene conc 1X 5.5 '
Milli Q water0.10.55
Takara HS Buffer5X211
dNTP2.5uM0.84.4
Primer Mix5uM4
HS Taq5U0.10.55
TemplateInstagene316.5
336.0ul / RXN
add 4.0ul Primer
A PCR cycle
98C10 sec
60C1 min
x35
AMP LaneTemplatePrimerAmt of Template
13 InstageneB13
23 InstageneC13
33 InstageneD13
43 InstageneE13
53 InstageneF13
64 InstageneB13
74 InstageneC13
84 InstageneD13
94 InstageneE13
104 InstageneF13
11AV 1ngB11
12AV 1ngC11
13AV 1ngD11
14AV 1ngE11
15AV 1ngF11
*AV 2ng Positive Control

Load 2.0ul Sample

Image:06may10B.jpg

AV 2ng - bad dilution

Nuclear Region HS Takara PCR Optimization

Compare 1.0ul of 20uM Primer vs 4.0ul of 5uM (old primer dilution)

HS Takara RXN Mixture conc 1X 3 '
Milli Q water0.10.3
Takara HS Buffer5X26
dNTP2.5uM0.82.4
Primer Mix5uM4
HS Taq5U0.10.3
TemplateInstagene39
186.0ul / RXN
add 4.0ul Primer
HS Takara RXN Mixtureconc1X3
Milli Q water3.19.3
Takara HS Buffer5X26
dNTP2.5uM0.82.4
Primer Mix20uM1
HS Taq5U0.10.3
TemplateInstagene39
279.0ul / RXN
add 1.0ul Primer
A PCR cycle
98C10 sec
60C1 min
x35
AMP LaneTemplatePrimerAmt of TemplatePrimer
13 InstageneF234.0ul of 5uM
24 InstageneF234.0ul of 5uM
3AV 20ngF214.0ul of 5uM
43 InstageneF231.0ul of 20uM
54 InstageneF231.0ul of 20uM
6AV 20ngF211.0ul of 20uM
*AV 20ng Positive Control

Load 2.0ul Sample

Image:07May10B.jpg

well 2 has a weak band

Personal tools