Team:Hawaii/Protocols/oligo hybridization

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
 * 1) Incubate at 37C for 1.5 hours.
 * 2) Add 4 μl 0.5 M NaCl.
 * 3) 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. | Oligonucleotide Inserts.