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 CCA3 1ng/ul 1 11
TopTaq buffer 10x 1 11
dNTP mix 2.5 mM each 0.2 2.2
Primer mix 5 uM each 0.5
TopTaq (QIAGEN) 5U/ul 0.05 0.55
Milli Q water 7.25 79.75
10 ul
PCR cycle
94C 3 min
98C 10 sec
60C 3 min
x35
72C 5 min
AMP Lane Contents Fosmid
1 AV10081 AV_TEL_L_A
2 AV10086 AV_TEL_L_A
3 AV10091 AV_TEL_L_A
4 AV10092.2 AV_TEL_L_A
5 AV10093.1 AV_TEL_L_A
6 AV10093.2 AV_TEL_L_A
7 AV100100 AV_TEL_L_A
8 AV10106 AV_TEL_L_B
9 AV10108 AV_TEL_L_B
10 AV10111 AV_TEL_L_B

Load 1.0ul sample 08march10 screen.jpg


Reaction mixture conc. 1x x4.5 ' PCR cycle (2 STEP) '
Temp (AV, CC A3) ?? 2 98C 10 sec
PrimeSTAR Buffer 5x 2 9 63C 1 min
dNTP mix 2.5 mM each 0.8 3.6 x35
Primer mix (AV10027) 5 uM each 0.5 2.25
PrimeSTAR HS DNA Polymerase 2.5 U/ul 0.05 0.225 PCR cycle (3 STEP)
Milli Q 4.65 20.925 98C 10 sec
10 ul 60C 5 sec
72C 1 min
x35

8Mar'10 TaKaRa-PS-Taq test.jpg

Alien Genes LongPCR

Amplification of AV240_B

' [CONC] 1X 4 '
Water 5.3 21.2
Takara LA Taq 5U 0.1 0.4
Takara LA Buffer 10X 1 4
dNTP 2.5uM each 1.6 6.4
*Template 1ng 1 4
Primer Mix 5uM each 1
10.0ul 36 9.0ul/RXN
add 1.0 Primer Mix
* AV or CCA3
PCR Cycle
94C 1 min
98C 10 sec
68C 15 min
30X
AMP Lane Template Contents ' Fosmid MIX
1 AV AV10029 FWD AV10034 RVS AV240_B 34
2 AV AV10032 FWD AV10040.1 RVS AV240_B 35
3 AV AV10040.1 FWD AV10041 FWD AV240_B 36
4 AV AV10010.1 FWD AV10042 FWD AV240_B 37
5 CCA3 AV10029 FWD AV10034 RVS AV240_B 34
6 CCA3 AV10032 FWD AV10040.1 RVS AV240_B 35
7 CCA3 AV10040.1 FWD AV10041 FWD AV240_B 36
8 CCA3 AV10010.1 FWD AV10042 FWD AV240_B 37

Load 1.0ul sample 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℃

8Mar'10 Covaris test.jpg