Team:Hawaii/Protocols/oligo hybridization
From 2008.igem.org
Hybridization of Parts
Protocol
- Mix:
- 3 μl 100 µM sense oligo
- 3 μl 100 µM anti-sense oligo
- 3 μl 10 x PNK (polynucleotide kinase) buffer
- 2 μl 10mM ATP
- 2 μl T4 polynucleotide kinase (PNK)
- 17 μl distilled water
- Total volume = 30 μl
- Incubate at 37C for 1.5 hours.
- Add 4 μl 0.5 M NaCl.
- Heated mixture in PCR machine to 97C for 2 min. Cooled mixture at each degree between 80 and 96 for 2 min (Tm range of oligos). Cooled mixture at each degree between 25 and 79 for 1 min.
- Reference: Pam Silver Lab. [http://openwetware.org/wiki/Silver:_Oligonucleotide_Inserts| Oligonucleotide Inserts.]
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