How do you reconstitute dry primers?

For every 1 nmoles, add 10 μL of PCR-grade water. For example, if a primer states 19.4 nmoles, then add 194 μL of PCR-grade water. Mix the solution by vortexing to reconstitute the primers.

How do you calculate primer dilution?

To determine the amount of water to add to the lyophilized primer simply multiply the number of nmol of primer in the tube by 10. That will be the amount of water to add to make a 100 µM primer stock. For example, if there are 38.2 nmol of primer a 100 µM primer stock is created by adding 382 µl of water.

How do you reconstitute primers Sigma?

Follow these steps:

1. Resuspend the oligonucleotide in 400 µL of water or buffer.
2. Dilute 12 µL into 988 µL of sterile, nuclease-free water.
3. Take an A260 reading of the 1 mL sample in a cuvette.
4. Ensure the OD value is in the linear range (~0.1 to 1 OD).
5. Multiply the OD of the sample volume by the dilution factor.

Is it OK to vortex primers?

Don’t vortex too much. But you can vortex gently for 5-10 for proper mixing. It would not break primers.

How do you do a 1/20 dilution?

For example, a 1:20 dilution converts to a 1/20 dilution factor. Multiply the final desired volume by the dilution factor to determine the needed volume of the stock solution. In our example, 30 mL x 1 ÷ 20 = 1.5 mL of stock solution.

Can you vortex oligonucleotides?

Most oligos are relatively easy to resuspend; however, those modified with fluorophores or hydrophobic molecules may require more time to become completely solubilized. If resuspension is difficult, try heating the oligo at 55°C for 1–5 minutes, then vortex thoroughly.

What is 1X TE buffer?

Description. This 1X TE Buffer is a component of the PureLink™ 96 Plasmid Purification System, now offered separately. It is used to resuspend the final purified plasmid pellet and contains very low EDTA, so it is compatible with sequencing and other enzymatic applications.

Can I Vortex DNTP?

Do not vortex PCR mix. Add DNA polymerase (Taq) to the reaction tube last. Avoid overloading PCR products into the gel; this may result in cross-contamination or misinterpretation of the results.