Rutgers:DNA Ligation

• Prepare a master-mix in a 0.5 ml tube containing (µl):

DW Tp2x pGEMT Ligase - for 1 tube: 1.5 2.5 0.25 0.5 (total: 4.75) - for 4 tubes: 6.0 10.0 1.0 2.0 (total: 19.0) - for 6 tubes: 9.0 15.0 1.5 3.0 (total: 28.5) - for 8 tubes: 12.0 20.0 2.0 4.0 (total: 38.0) - for 10 tubes: 15.0 25.0 2.5 3.0 (total: 47.5) - for 12 tubes: 18.0 30.0 3.0 6.0 (total: 57.0)

• dispense 4.75 l of the mix into 0.2 ml tubes
• Add 0.25 µl of the purified PCR-product
• vortex
• incubate 1 night at 4°C or 1 h at RT
• spin before use