Team:Glendale CC AZ/Protocols/Ligation

Glendale Community College Arizona

Protocols

Growth Curve Assay

NaCl Growth Curve Assay

Survival Growth Assay

Alkaline Lysis Plasmid Miniprep

Restriction Digest

DNA Isolation

Bioinformatics

Ligation

Transformation

Ligation Protocol

Protocol from: http://parts.igem.org/Help:Protocols/Ligation

Materials

  -digested plasmid backbone
  -equimolar amount digested Part 1
  -equimolar amount digested Part 2
  -T4 DNA ligase buffer
  -T4 DNA ligase
  -dH2O
  -thermal cycler

Procedure

1. Add 2µL of digested plasmid backbone

2. Add equimolar amount of digested Part 1

3. Add equimolar amount of digested Part 2

4. Add 1µL T4 DNA ligase buffer.

5. Add 0.5µL T4 DNA ligase.

6. Add water to 10µL.

7. Place in pre-programmed thermal cycler at 16°C for 30 minutes then at 80°C for 20 minutes.