User:Alexander S Mikheyev/Notebook/rotifer alien genes/2009/12/11
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Cycle Sequencing Clones M13FWD83 colonies PCR amped -> sequencing Nanodrop Data
|
Cycle Sequencing | ' | ' | ' |
Reagents | Concentration | 1X | 87X |
BDT | 0.5 | 43.5 | |
M13 FWD | 0.4 | 34.8 | |
DMSO | 0.1 | 8.7 | |
5X Seq Buffer | 5X | 0.4 | 34.8 |
H20 | 3.6 | 313.2 | |
PEG DNA | variable | 1 | |
TOTAL | 6 | 435 | |
Cocktail per well = 5.0ul | |||
PCR | 96C | 10 sec | |
50C | 5 sec | ||
60C | 4 min | ||
60 cycles | |||
Cycle Sequencing AV10007, AV10016, AV10027, AV10153, PR10008 and 5475F + 8047R
Cycle Sequencing | ' | ' | ' |
Reagents | Concentration | 1X | 14X |
BDT | 0.5 | 7 | |
DMSO | 0.1 | 1.4 | |
5X Seq Buffer | 5X | 0.4 | 5.6 |
H20 | 3.6 | 50.4 | |
*Primer | 15uM | 0.4 | |
*PEG DNA | variable | 1 | |
TOTAL | 6 | 64.4 | |
Cocktail per well = 4.6ul | |||
*add primer and PEG separately | |||
PCR | 96C | 10 sec | |
50C | 5 sec | ||
60C | 4 min | ||
60 cycles | |||
Primers | Sequence FWD and RVS | ||
AV10007 | |||
AV10016 | |||
AV10027 | |||
AV10153 | |||
PR10008 | |||
5475F + 8047R | |||