User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/27

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Contents

Cycle Sequence AMPure Cleaned PCR Product

HCO primer only

CS Reaction mixture conc. 1x 90 '
Big Dye Terminator0.545
Primer (FWD or RVS)5uM0.436
DMSO0.19
5X Seq Buffer5X0.436
Template *?4.6
1261.4ul / RXN
* PEG Cleaned PCR product
Cycle Seq Program
96C10 sec
50C5 sec
60C4 min
x60
CS Plate Source Plate Source Plate Well EX ID CS Plate Source Plate Source Plate Well EX ID
A1BaltimoreA11D6BaltimoreA344
B1BaltimoreB12E6BaltimoreA645
C1BaltimoreC23F6E. WindsorD446
D1BaltimoreC34G6HarvardA147
E1E. WindsorA15H6Ithaca 2A648
F1E. WindsorA36A7Pennsylvania (PA)D449
G1E. WindsorB17B7Tsukuba 1B550
H1E. WindsorB28C7YokohamaA251
A2E. WindsorC29D7BaltimoreC152
B2HarvardA210E7Ithaca 1A453
C2HarvardA311F7Pennsylvania (PA)B654
D2Ithaca 1A112G7YokohamaB255
E2Ithaca 1A213H7YokohamaA456
F2Ithaca 1A314A8Ithaca 1C457
G2Ithaca 1A515B8Ithaca 1D558
H2Ithaca 1A616C8BaltimoreC559
A3Ithaca 1B117D8E. WindsorA260
B3Ithaca 1B318E8Pennsylvania (PA)B361
C3Ithaca 1B519F8Ithaca 1C362
D3Ithaca 1C120G8Ithaca 1C663
E3Ithaca 1D121H8HarvardA464
F3Ithaca 1D322A9Ithaca 1B465
G3Ithaca 1D423B9Ithaca 1C566
H3Ithaca 1D624C9BaltimoreB267
A4Ithaca 2A125D9BaltimoreB368
B4Ithaca 2A226E9BaltimoreB669
C4Ithaca 2A327F9E. WindsorC370
D4Ithaca 2A428G9Pennsylvania (PA)D671
E4Ithaca 2A529H9BaltimoreB572
F4Pennsylvania (PA)B430A10Ithaca 1C273
G4Tsukuba 1A431B10Pennsylvania (PA)D574
H4Tsukuba 1A632C10Woods Hole 1B3E
A5Tsukuba 1B233D10Woods Hole 1B5F
B5Tsukuba 1B434E10Woods Hole 1C4G
C5Tsukuba 1C135F10Woods Hole 2A6H
D5Tsukuba 1C636G10Cape Cod (CC)A3CCA3
E5Tsukuba 3D237H10BostonA1A
F5Tsukuba 3D338A11BostonA4B
G5Tsukuba 3D439B11Cape Cod (CC)A1C
H5Woods Hole 1A640C11Woods Hole 3A5D
A6Woods Hole 1C241D11Woods Hole 3C2I
B6Woods Hole 1C342E11BaltimoreA575
C6YokohamaB543F11Pennsylvania (PA)B176

Nuclear Gene Rotifer Culture 454 Amplification

Adenita Culture 1-12

EX Takara RXN Mixture conc 1X 6.5 '
Milli Q water5.1533.475
Takara EX Buffer10X16.5
dNTP2.5uM0.85.2
Primer Mix20uM1
EX Taq5U0.050.325
TemplateInstagene213
58.59.0ul / RXN
A PCR cycle
98C10 sec
60C1 min
x35
Plate 1
Source PlateWellEX ID
BaltimoreA11
BaltimoreB12
BaltimoreC23
BaltimoreC34
E. WindsorA15
E. WindsorA36
E. WindsorB17
E. WindsorB28
E. WindsorC29
HarvardA210
HarvardA311
Ithaca 1A112
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