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Team:TU Darmstadt/Protocols/
From 2012.igem.org
(Difference between revisions)
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'''References''' | '''References''' | ||
| - | Lund, A. H., M. Duch, et al. (1996). "Increased cloning efficiency by temperature-cycle ligation." Nucleic Acids Res 24(4): 800-801. | + | *Lund, A. H., M. Duch, et al. (1996). "Increased cloning efficiency by temperature-cycle ligation." Nucleic Acids Res 24(4): 800-801. |
| - | Der Experimentator: Molekularbiologie / Genomics/6.Auflage/2009/C.Mülhardt | + | *Der Experimentator: Molekularbiologie / Genomics/6.Auflage/2009/C.Mülhardt |
Latest revision as of 21:41, 26 September 2012
Ligation reaction mix
- 10x NEB 4 Buffer 2µL
- T4 Ligase 1µL
- Vector 50 ng
- Insert X
- H2O ad 20µL
The insert to vector molar ration of 1:5 was chosen and caltulated with the formula:
A ligation with T4 DNA Ligase(purchased from NEB) followed.
Schmidt´sche thermocycler ligation program (STLP)
- T= 4.0°C 10 min
- T= 10°C 60 min
- T= 12°C 15 min
- T= 16°C 10 min
- T= 25°C 10 min
- T= 37°C 5 min
- T= 70°C 20 min
References
- Lund, A. H., M. Duch, et al. (1996). "Increased cloning efficiency by temperature-cycle ligation." Nucleic Acids Res 24(4): 800-801.
- Der Experimentator: Molekularbiologie / Genomics/6.Auflage/2009/C.Mülhardt
