User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/14

From OpenWetWare
Jump to: navigation, search
OpenWetWare photo.jpg Alien genes Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Nuclear Gene

Amplification of Adenita Cultures

Reaction mixture conc. 1x 6.5 '
Template Instagene ? 1 6.5
TopTaq buffer 10x 1 6.5
dNTP mix 10 mM each 0.2 1.3
Primer mix 5 uM each 1
TopTaq (QIAGEN) 5U/ul 0.05 0.325
Milli Q water 6.75 43.875
10 ul 58.5 9.0ul / RXN
PCR cycle
95C 3 min
95C 5 sec
60C 1 min
x35
AMP Lane Template Primer
1 71 A1
2 71 A2
3 71 A3
4 71 A4
5 71 A5
6 71 A6
7 72 B1
8 72 B2
9 72 B3
10 72 B4
11 72 B5
12 72 B6

File:14April10.tif

Nuclear Gene

Troubleshoot PCR Program

Reaction mixture conc. 1x 4 '
Template AV 20ng/ul 1 4
TopTaq buffer 10x 1 4
dNTP mix 10 mM each 0.2 0.8
Primer mix 5 uM each 1
TopTaq (QIAGEN) 5U/ul 0.05 0.2
Milli Q water 6.75 27
10 ul 36 9.0ul / RXN
A PCR cycle
95C 3 min
95C 5 sec
60C 1 min
x35
B PCR cycle
95C 3 min
95C 5 sec
60C 1 min
x10
95C 5 sec
68C 1 min
x25
AMP Lane Template Primer PCR Program
1 AV A1 A
2 AV *AV10027 A
3 AV A1 B
4 AV *AV10027 B
* AV10027 Positive Control

Load 4.0ul of sample

File:15April10.tif

Genome extraction

Tsukuba3 D4(QIAamp DNA micro kit)
100ul EB (3.4 ug/ul)

load 1.0ul of extraction

0076.jpg


Alien Genes New Primer Screen

Reaction mixture conc. 1x x33 ' Lane Gene Lane Gene
Temp (Tsukuba3 D4) 1ug/ul 1 33 1 AV10158 17 AV10084
TopTaq Buffer 10x 1 33 2 AV10153 18 AV10082
dNTP mix 10 mM each 0.2 6.6 3 AV10151 19 AV10075
Primer mix 5 uM each 1 4 AV10150 20 AV10071
TopTaq 5 U/ul 0.05 1.65 5 AV10140 21 AV10045
Milli Q 6.75 222.75 6 AV10134 22 AV10043
10 ul 7 AV10130 23 AV10042
8 AV10123 24 AV10041
PCR cycle 9 AV10121 25 AV10034
94C 3 min 10 AV10104 26 AV10028
11 AV10109 27 AV10027
98C 10 sec 12 AV10099 28 AV10025
60C 3 min 13 AV10098 29 AV10016
x35 14 AV10094 30 AV10011
15 AV10092 31 AV10007
72C 5 min 16 AV10089 32 AV10002

load 2.5ul of sampple

0079.jpg