User:Anthony Salvagno/Notebook/Research/2009/09/02/Sap Cap AGC suspension and annealing

For protocol see here: Resuspension of T, B, and HP oligos

Resuspension
pmols:17718
 * 1) add 177uL of .1x TE
 * 2) After addition of liquid I vortex the solution for about 1 min.
 * 3) Let the stocks sit on ice for 60'
 * 4) Vortex for 1min again.
 * 5) Take nanodrop reading:
 * 6) *820ng/uL - with a MW of 8929g/mol this gives me a concentration of 92uM which is close enough!
 * 7) *again, we want 100uM

Annealing
The idea is to heat and quick chill. So I am going to follow the protocol on above mentioned page: 5min at 100C and 5min on ice.

I really don't want to make a google doc for this one reaction so I'll just detail below:
 * 1) made annealing buffer from 10x TE and 3.8M NaCl:
 * 2) *1uL of 3.8M NaCl in 75uL 10x TE (that is my logic to get 50mM NaCl + TE)
 * 3) combined
 * 4) *6uL sapcap
 * 5) *10uL 10x anneal buff
 * 6) *84uL ddH2O
 * 7) heat at 95C for 5 min
 * 8) cool on ice for 5 min
 * 9) store in box #3 as Sap Cap

Tomorrow is ligation day.

Basing off the above results, I put 5ug DNA in the above reaction. That will give me 50ng/uL in solution for a total of 5.6uM. I don't know if making it double stranded will decrease the molarity or what. I could do a nanodrop, but I don't wanna.