Daniel:Notebook/ComboLock/2017-6-16
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Antibody Detection Test-Positive Control (Started Wednesday)
Protocol-Part 5
- Bglii digestion
- Make the following reactions; Do not add Bglii yet
- Aliquot 12 uL master mix into new tube
- Add 7 uL appropriate sample
- Mix with vortexer, spin down, and add 1 uL Bglii
- Incubate 2 hr 37C
- Heat kill with 65C for 20 min
- Hold at 12C
Reagent | Single Rxn Vol | Master Mix (6.2X) |
RCA Rxn | 7 | 0 |
10X Buffer 3.1 | 2 | 12.4 |
Bglii | 1 | 6.2 |
nfH2O | 10 | 62 |
Total | 20 | 80.6 |
- Make the following 7.2X master mix (1X)
- 7.2 uL 10 uM AmpF6.4Sol
- 151.2 uL (21) nfH2O
- 180 uL 2X Kapa SYBR Fast Master Mix
- Add 47 uL master mix to each lane
- Add 1 uL AmpR and 2 uL sample according to plate layout
- Use the following protocol