"
Common Laboratory Protocols
- Bacterial Transformation
- Colony PCR
- Gel Extraction
- Miniprep
- PCR Purification
- PCR Reaction
- Restriction Enzyme Digest
- TdT Heat Inactivationn
- TdT Nucleotide Addition
- Yeast Gene Knockout
- Yeast Mating
- Yeast Sporulation/Zymolyase Treatment
Bacterial Transformation
Colony PCR
Gel Extraction
Miniprep
PCR Purification
PCR Reaction
Restriction Enzyme Digest
- Materials
- 10x Buffer (Compatible to the enzyme)
- Enzyme
- BSA - diluted to 10x
- ddH2O
- DNA - up to 1μg per reaction.
Procedure Verify all times and temperatures for specific enzyme prior to use. This is general protocol.
- Prewarm 2 water baths to 37°C and 70°C
- Thaw buffers, diluted BSA. Keep all reagents on ice
- Set up reactions based on table below.
Reagent Volume Example Reaction Buffer 1/10 total volume 2μL Diluted BSA (10x) 1/10 total volume 2μL DNA calculate for target ng 3.7μL ddH2O volume up to (total volume - 1μL) 11.3μL Enzyme generally 1μL 1μL Total Volume 20uL - Incubate 60min 37°C.
- Heat inactive 70°C for 15 min.
- Store in -20°C or use immediately.
Top of Page
