What concentration should my primer be?
All primers should be sent at 10µM.
What volume of primer should I send?
- Primers should be provided at a concentration of 10µM (picomoles/µl).
- For 96-well format, provide at least 50 µl of primer in a tube for each full plate.
- For primer plates, provide at least 6ul of primer per well of the plate.
- For individual samples, provide 2 µl of primer per sequencing reaction + 5 µl.
- Example: 4 sequencing reactions = 8 µl + 5 = 13 µl total primer to send.
Can I premix the sample and primer?
Yes. Be sure you have a total of 20 picomoles of primer per 15µl of premix. We recommend using the following formula to create pre-mix samples:
DNA: N=0.1 x (vector size in base pairs + insert size in base pairs)
*N is the nanograms of DNA that should be added to each 15µl of premix.
What should the sample concentration be?
The sample type will determine the concentration needed.
i. PCR Product: 25-50 ng/µl
ii. Plasmid DNA: minimum of 50 ng/µl but we prefer it at 100 ng/µl. If you are sending it at 100ng/µl we need 10µl for one reaction or 15µl for two reactions. If you are sending it at less than 100ng/µl please send 15µl for one reaction or 25µl for two reactions.
*Note: If your samples are below the desired concentrations as listed above, we can still sequence them, the results are just less likely to be of high-quality. Please indicate the concentration on your order form if known so that we can adjust the sample volume to add per reaction accordingly.
I am sending cultured pellets and am unsure if they are sufficient. Do you have any suggestions?
If you can check the O.D. of the solution before pelleting, it should be around 3.0-4.0. If you did not check the O.D. prior to pelleting , feel free to send the pellets and, if upon arrival they appear too small, we will contact you and inquire about re-growing your samples here at our facility if needed.
*If we have to re-grow them this will add an additional day for turnaround time.
Do I have to send my own primer?
No. We have a list of free primers. We can also synthesize and/or design your primers for you. Primers are stored for a period of 2 years.
What primers do you offer for free?
Please refer to the primers page on the website for a complete list.
Can you do primer synthesis?
Yes. It is $16 per primer for standard primers. If you want us to design and synthesize the primer the cost is $25 per primer.
Can you quantify my samples?
Yes, if you would like any of your samples quantified, please indicate which samples you would like us to quantify in the notes section of your order form. We CANNOT quantify crude PCR products, or premixed samples.