DNA Probes Resuspension

Calculating resuspension:

We want to make a stock solution of 100mM solution when resuspending. Take the amount in NMOL given in the QC sheet with the primers. Take that value and multiply by 10. This will give you the amount in mL of TE to add to the well. The calculation is as follows assuming the amount ordered is close to 10nmol

~10 x 10^-9 mol = 0.1 x 10^-3 mol/L = 100mM

100 x 10^-6 L

Reconstitution and Storage:

To reconstitute: Centrifuge the tube for a few seconds to collect the DNA in the bottom of the tube. Carefully open, add an appropriate volume of TE buffer, close, rehydrate (let stand) for two minutes, and vortex for 15 seconds. It is recommended that primers be reconstituted at concentrations greater than 10 µmolar in TE (10mM Tris-HCl, pH 8.0, 1mM EDTA) and stored at -20°C. Stability (when kept at -20°C) of lyophilized material is >1 year and the stability of reconstituted material is >6 months. Spin again briefly to collect at the bottom.

* Note: HEX, TET, FAM, fluorescein and rhodamine-labeled primers are sensitive to light. Exposure of these oligonucleotides is kept to a minimum during synthesis and shipping. Upon receipt, please store in the dark. Storage in a black box is recommended.

Preparation of Working Stock Solution:

From the 100mM stock solution make the 2mM working stock solution as follows. In a 96 well plate, aliquot 96mL of ddH₂O to each well. Add 2mL of the forward primer and 2mL of the reverse primer in to the same well using the 8-channel pipetter. This will give a final concentration of 2mM of both the forward and reverse primers mixture. Place a plate cover over the plate and store at -20°C.