AlanFung:LabNotes/Probe/2013-12-11: Difference between revisions
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*Molar Mass of ssDNA probe: 146bp * (325Da/bp) = 146bp * (325 g/mol/bp) = 47450 g/mol | *Molar Mass of ssDNA probe: 146bp * (325Da/bp) = 146bp * (325 g/mol/bp) = 47450 g/mol | ||
*(32.71ng/ul * 1*10^6 ul/L) / (47450g/mol) = 689.36nM | *(32.71ng/ul * 1*10^6 ul/L) / (47450g/mol) = 689.36nM | ||
*I will use 100nM final concentration for the expansion PCR | |||
===GP1_V4 Expansion PCR=== | ===GP1_V4 Expansion PCR=== | ||
===GP1_V6 Expansion PCR=== | ===GP1_V6 Expansion PCR=== |
Revision as of 00:09, 12 December 2013
GP1 Padlock Probe Production
Overview
- Expansion PCR
- Production PCR
- Exonuclease digestion to generate SS-DNA
- Removal of amplification adapters
- Probe purification by PAGE size-selection
Expansion PCR
- Probe concentration from CustomArray 32.71ng/ul volume 80uL Amount 2.62ug
- Probe size 146bp
Convert ng/ul to nM
- Molar Mass of ssDNA probe: 146bp * (325Da/bp) = 146bp * (325 g/mol/bp) = 47450 g/mol
- (32.71ng/ul * 1*10^6 ul/L) / (47450g/mol) = 689.36nM
- I will use 100nM final concentration for the expansion PCR