November
29
Lab Notebook: 11/21 Restriction Enzyme Digests
Name: Roshni Jaffery
Date: 11/21/16
Lab: Bacteriophage Lab – Day 26
Results: I extracted my phage’s DNA in my last lab and we measured my nano drop which was 58.5 ng/µl.
Goal: To cut DNA into fragments based on the DNA’s sequence.
Materials:
- Phage DNA
- Restriction Enzymes with buffers
- microcentrifuge tubes
- water bath/ incubator
- heat block
Procedure:
- Flick the DNA sample to prepare it.
- Incubate the DNA at 65 degrees Celsius for 10 minutes and quickly place it on ice.
- Micro centrifuge for less than a minute.
- Set up reaction in each of the micro centrifuge tubes as shown in the table below.
- Micro centrifuge each tube for less than a minute.
- Incubate all the micro centrifuge tubes at 37 degrees Celsius for an hour.
- Store in the freezer.
| Centrifuge 1 | Centrifuge 2 | Centrifuge 3 | Centrifuge 4 | Centrifuge 5 | |
| (NOTE: units in µl) | Uncut | BstE | Scal | EcoR1 | HindIII |
| DNA | 9 | 9 | 9 | 9 | 9 |
| Enzyme | 0 | 2.5 | 2.5 | 1 | 1 |
| 10x EcoR1 Buffer | 2.5 | – | – | 2.5 | 2.5 |
| Water | 13.5 | 13.5 | 13.5 | 12.5 | 12.5 |
Labels: Uncut or enzyme used, RJ, 11/21 @ freezer
Next Steps: Perform a gel electrophoresis.