Dinh/NOTES/2010-10-21
Jump to navigation
Jump to search
Prepare DMR220K Suppressor Oligos[edit]
- Typical DMR220K capture reaction uses 200ng gDNA, which is 1.10E-019 moles template
- Moles of probe, 100:1 ratio, required is 1.10E-017 moles probe
- Use 100:1 suppressor oligo to probe ratio is 1.10E-015 moles suppressor oligo
- Will use 1ul of suppressor oligo mixture per capture reaction, which means concentration of suppressor oligo should be 1.1nM
- We have 96 suppressor oligo = 1.1nM*96 = 105nM
Dilute all 96 to 1.05uM and then combine all to obtain a final mixture of 1.05uM of suppressor oligo mixed.[edit]
- DMR220Ksuppressoroligo from IDTDNA is at 10uM concentration
- Add 1ul of 10uM oligo to 8.53ul of water to get 1.05uM
- 8.53x96 = 818.29ul water
- In a solution basin, add 8.53ulx96 = 818.29 ul of ddH2O.
- Use Rainin Multichannel p10 to add 1ul of each 10uM suppressor oligo to solution basin.
- Finally, shake/ use P1000 to mix well, and pipette the mixture into a labeled 1.5mL tube.