User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/03/08

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Alien Genes New Primer Screen

Reaction mixture conc. 1x 11
Template CCA31ng/ul111
TopTaq buffer10x111
dNTP mix2.5 mM each0.22.2
Primer mix5 uM each0.5
TopTaq (QIAGEN)5U/ul0.050.55
Milli Q water7.2579.75
10 ul
PCR cycle
94C3 min
98C10 sec
60C3 min
x35
72C5 min
AMP Lane Contents Fosmid
1AV10081AV_TEL_L_A
2AV10086AV_TEL_L_A
3AV10091AV_TEL_L_A
4AV10092.2AV_TEL_L_A
5AV10093.1AV_TEL_L_A
6AV10093.2AV_TEL_L_A
7AV100100AV_TEL_L_A
8AV10106AV_TEL_L_B
9AV10108AV_TEL_L_B
10AV10111AV_TEL_L_B

Load 1.0ul sample Image:  08march10 screen.jpg


Reaction mixture conc. 1x x4.5 ' PCR cycle (2 STEP) '
Temp (AV, CC A3)??298C10 sec
PrimeSTAR Buffer5x2963C1 min
dNTP mix2.5 mM each0.83.6x35
Primer mix (AV10027)5 uM each0.52.25
PrimeSTAR HS DNA Polymerase2.5 U/ul0.050.225PCR cycle (3 STEP)
Milli Q4.6520.92598C10 sec
10 ul60C5 sec
72C1 min
x35

image: 8Mar'10_TaKaRa-PS-Taq_test.jpg

Alien Genes LongPCR

Amplification of AV240_B

' [CONC] 1X 4 '
Water5.321.2
Takara LA Taq5U0.10.4
Takara LA Buffer10X14
dNTP2.5uM each1.66.4
*Template1ng14
Primer Mix5uM each1
10.0ul369.0ul/RXN
add 1.0 Primer Mix
* AV or CCA3
PCR Cycle
94C 1 min
98C10 sec
68C15 min
30X
AMP Lane Template Contents ' Fosmid MIX
1AVAV10029 FWDAV10034 RVSAV240_B34
2AVAV10032 FWDAV10040.1 RVSAV240_B35
3AVAV10040.1 FWDAV10041 FWDAV240_B36
4AVAV10010.1 FWDAV10042 FWDAV240_B37
5CCA3AV10029 FWDAV10034 RVSAV240_B34
6CCA3AV10032 FWDAV10040.1 RVSAV240_B35
7CCA3AV10040.1 FWDAV10041 FWDAV240_B36
8CCA3AV10010.1 FWDAV10042 FWDAV240_B37

Load 1.0ul sample Image:  08march10 LongPCR.jpg

Apis mellifera methylome

DNA sample (callow worker) 2.5 ug / 100 ul
Fragmentation by Covaris
・Program:
Target Base Pair (Peak) 300bp
Duty Cycle 10%
Intensity 4
Cycle per Burst 200
Time (second) 120
Chiller Temperature 19.5℃

image: 8Mar'10_Covaris_test.jpg

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