Primer Walking & Plasmid Sequence Confirmation Minimum and Recommended Sample Requirements
|
Sample Concentrations |
|
Plasmids |
PCR Products |
With ITRs or difficult regions* |
Minimum |
100 ng/µl |
10 ng/µl |
200 ng/µl |
Recommended |
200 ng/µl |
20 ng/µl |
300 ng/µl |
|
Sample Volumes |
|
Plasmids |
PCR Products |
With ITRs or difficult regions* |
|
1x coverage |
2x coverage |
4x coverage |
1x coverage |
2x coverage |
4x coverage |
1x coverage |
2x coverage |
4x coverage |
Minimum |
5 µl per kb |
6 µl per kb |
12 µl per kb |
5 µl per kb |
6 µl per kb |
12 µl per kb |
+ 4 µl per region |
+ 4 µl per region |
+ 8 µl per region |
Recommended |
10 µl per kb |
12 µl per kb |
24 µl per kb |
10 µl per kb |
12 µl per kb |
24 µl per kb |
+ 8 µl per region |
+ 8 µl per region |
+ 16 µl per region |
* Please include this amount in addition to the standard plasmid and PCR product volumes. Difficult regions include large repeats, G-C rich regions, homopolymeric regions (e.g. CAAAAAAA), etc.
Note: The received sample volume must be at least 5 µl in order to process.
Primers:
Our sequencing experts at Azenta will use proprietary primer design to create and synthesize primers based on the provided reference sequence (or known sequence) for each sample. If you would like to send your own primer aliquots for the initial round of sequencing, please submit a minimum of 3 µl per sample at a concentration of 5 µM. If submitting fewer than 4 samples, please use a minimum primer volume of 10 µl.
Packaging:
Please send samples/primers in one of the following formats:
- 1.5 – 2.0 ml screw cap or flip-top microcentrifuge tubes
- 8-strip PCR tubes
Please secure all sample lids to prevent spilling and contamination during transit.
You may submit at ambient temperature or with blue ice.
Please print and include your order confirmation page, which contains a barcode with the order tracking #.
Shipping:
You may ship your samples using a standard overnight service to the address listed below, or you may use an Azenta Life Sciences Drop-box/courier service if one is available near you.