Baylor-exon-Sept09: Difference between revisions
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*Note: Amplification started after ~13 cycles | *Note: Amplification started after ~13 cycles | ||
'''Ethanol Precipitation''' | |||
Collected 24 reactions in a 16mL tube and added: | |||
0.1x 3M NaOAc (240ul) | |||
2.5x 100% EtOH (6mL) | |||
8ul GlycoBlue | |||
-> Allow to cool in -70C freezer for at least 20 minutes | |||
-> Spin in 4C at 4k rpm for 20min | |||
-> Remove all liquid | |||
-> Suspend palette in 750ul 75% EtOH | |||
-> Move solution from 16mL tube to fresh 1.6mL tube | |||
-> Spin in 4C at 10k rpm for 5min | |||
-> Remove all liquid, allow palette to dry | |||
-> Suspend palette in 100ul H2O |
Revision as of 02:24, 4 November 2009
Baylor (BCM) Creation
PCR
1x 48x Template 0.4ul 19.2ul 2x Kapa Master Mix 50ul 50ul H2O 50ul 50ul eMIP-CA1F (100uM) 0.4ul 0.4ul eMIP-CA1R (100uM) 0.4ul 0.4ul 95C, 30sec -> (95C, 3sec -> 54C, 30sec -> 60C, 20sec -> plate read) x 25 -> 60C, 2min -> 15C hold
- Note: Amplification started after ~13 cycles
Ethanol Precipitation
Collected 24 reactions in a 16mL tube and added: 0.1x 3M NaOAc (240ul) 2.5x 100% EtOH (6mL) 8ul GlycoBlue -> Allow to cool in -70C freezer for at least 20 minutes -> Spin in 4C at 4k rpm for 20min -> Remove all liquid -> Suspend palette in 750ul 75% EtOH -> Move solution from 16mL tube to fresh 1.6mL tube -> Spin in 4C at 10k rpm for 5min -> Remove all liquid, allow palette to dry -> Suspend palette in 100ul H2O