Daniel:Notebook/ComboLock/2017-3-13
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Bolt Test (Started Wednesday March 8)
Protocol
Sample Matrix
Sample | Condition |
Sample 5 (AB) | No Ligase |
Sample 6 (AB) | Normal |
- Template-Bead Binding
- Suspend 2 uL (8 ug) beads per sample in 100 uL wash buffer
- Apply magnet for 30 sec and remove supernatant
- Add 2 uL 10 uM template oligo per sample to bead solution; incubate at RT for 5 min
- Wash beads with 100 uL wash buffer; vortex to suspend; apply magnet and remove supernatant
- C Probe Hybridization
- Combine following into a 0.2 mL tube
- Heat C probes to 90C for 5 min; Chill probes on ice to quench
- Add 5 uL probe mixture to beads
- Incubate at 40C for 1 hour with agitation
- Wash beads with 100 uL wash buffer; apply magnet and remove supernatant
- Repeat wash step above
- Repeat wash step above
Reagent | uL per Sample | Exp uL Total | No C uL Total | NP uL Total |
PCCA-03 (10 uM) | 1 | 4 | 0 | 2 |
PCCB-03 (10 uM) | 1 | 4 | 0 | 0 |
Mirror BC-v2 (10 uM) | 1 | 0 | 0 | 2 |
Wash buffer | 3 | 12 | 10 | 6 |
Total | 5 | 20 | 10 | 10 |