Daniel:Notebook/ComboLock/2017-7-8
Jump to navigation
Jump to search
Oligo+Protein Test (Started Thursday)[edit]
Protocol[edit]
- qPCR
- Make 9.2X master mix according to following (1X)
- 193.2 uL nfH2O (21)
- 9.2 uL 10 uM AmpF6.4Sol (1)
- 230 uL 2X Kapa SYBR Fast Master Mix (25)
- Add 47 uL master mix to each well
- Add 2 uL sample and 1 uL AmpR Index (10 uM) according to plate layout File:PlateLayout-20170708-Ab-and-Oligo.png
- Run the Sstandard qPCR protocol for 23 cycles
- TBE Gel
- Mix together 120 uL TBE and 30 uL 6X dye
- Aliquot 10 uL/lane onto parafilm
- Add 2 uL correct sample or (1.5 uL for ladder) to aliquots
- Add 10 uL aliquots to correct lanes
- Run gel at 235V for 24 minutes
- Remove gel from case and incubate in TBE buffer with 2 uL SYBR Gold for 3 minutes
- Image in gel doc
Results[edit]
- PlateLayout-20170708-Ab-and-Oligo.png
Plate layout
- Ab-and-Oligo-20170708-plateCTs-normed.png
Plate CTs
- Ab-and-Oligo-20170708-rawcurves.png
Raw curves
- 2017-07-08-Oligo-and-Ab.png
Gel image