LAB - GENE CLONING

Day 0:

• Prepare bacteria E. coli strains (Blue, Red, GFP)

• Prepare primers for PCR

• Prepare plasmid extraction kit

• Prepare PCR cleanup kit

• Prepare PCR kit

• Prepare restriction enzyme kit: EcoRI, PstI

• Prepare ligation kit: ligase

• Prepare agarose gels for DNA electrophoresis and the related instrument

• Prepare pipettes

Day 1: DNA --> PCR --> RE cut

1. Pipette usage

2. Plasmid extraction

3. DNA concentration measurement

4. PCR

5. ------ LUNCH BREAK ------

6. Gel electrophoresis

7. PCR cleanup

8. Restriction enzyme digestion

Day 2: DNA ligation --> Transformation

1. Cleanup

2. Gel electrophoresis

3. Ligation

4. ------ LUNCH BREAK ------

5. Transformation

6. Bacterial culture

Day 3: Observe the resulting colonies

• see if GFP-positive colonies grow