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.

Ship to:

ATTN: Primer Walking
Azenta Life Sciences
Hope End, Takeley
Essex CM22 6TA

Use an Azenta Drop-box/Courier

Contact us to set up or inquire about a drop-box near you!
Primer.Walking@Azenta.com

If you have any questions or concerns regarding sample requirements, please contact us. Our team is happy to help you!