Richard Lab:Restriction Digest protocol
- prepared DNA (from Miniprep, PCR or Gel Extraction)
- restriction endonucleases
- 10X restriction endonuclease buffer
- BSA (optional)
- phosphatase buffer
- distilled water
- Sterile 1.5-ml microcentrifuge tubes
- Quickly vortex all ingredients (Buffer, BSA, DNA, Enzymes) before beginning.
- Measure out prepared DNA into sterile 1.5-ml microcentrifuge tube (1).
Add 4 volumes distilled water.
Use the following table as a checklist for preparing the reaction in sterile 1.5-ml microcentrifuge tube (2):
10X restriction endonuclease buffer BSA DNA solution restriction endonucleases Restriction Digestion 5 µl 1 µl 40 µl 1 µl