User:Anthony Salvagno/Notebook/Research/2009/04/27/Oligo Annealing

So last week sometime we got the oligos we ordered. One is the top oligo for the SapI adapter, and the other is a hairpin adapter with a NotI site. I need to resuspend the DNA and then make stocks.

Resuspension
I want 100uM stocks (100pmol/uL). I started with:
 * Top adapter: 11504pmol
 * Hairpin adapter: 27590pmol


 * 1) So I need to mix:
 * 2) *115uL 0.1x TE into TA
 * 3) *276uL 0.1x TE into HP
 * 4) After addition of liquid I vortex the solution for about 1 min.
 * 5) Let the stocks sit on ice for 60'
 * 6) Vortex for 1min again.
 * 7) Take nanodrop reading

Nanodrop results:
 * Top Oligo: 554ng/uL
 * Bottom Oligo: 2258.5ng/uL
 * Hairpin Oligo: 2442.6ng/uL

This yields: based on molecular weight.
 * Top: 45uM
 * Bottom: 217uM
 * HP: 84uM

Annealing

 * 1) Mix solutions to above specifications.
 * 2) Let both tubes sit for 5min at 100C.
 * 3) To cool:
 * 4) *Tube HP (hairpin) must sit in ice bath for 5min
 * 5) *Tube T+B (Top and Bottom oligos) must sit in block and cool to room temp.