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DNA-sequence

Waikato DNA Sequencing Facility (WDSF)

Sample Preparation

When preparing samples for submission to the WDSF, please first download our request form, complete and print it, ensuring it is included with your templates upon submission.

Please refer to the below requirements for each the different types of services we offer. Please also make sure to refer to both the troubleshooting section and to our disclaimer.

Find out more about results and archiving.



Template Requirements

  • The quality of any sequence or genotype is directly proportional to the quality of the template DNA, the chosen primer and their effective interaction. Samples and primers should ideally be resuspended in high grade water or suitable low pI buffer which must contain less than 0.1mM of any chelating agent.

    For example: 1x TE is not a suitable elution buffer for use with template or primers destined for DNA sequencing as it contains more than 0.1mM of EDTA.

  • Please note that many commercially available kits use similar levels of chelating agents in their elution buffers. In many cases this can be successfully substituted with a simple Tris based buffer.
  • Samples and Primers should be supplied free of residual salts, solvents, proteins, primers, RNA, chaotropic or chelating agents.
   Double Stranded 
DNA
 Single Stranded 
DNA
 PCR Products 

 Recommended
  concentration

150 ng/µL 50 ng/µL 10-45 ng/µL

 

  • For PCR products, the amount of DNA used in each reaction is dependant on product length. As a guide this is usually 3 ng per 100 bp (template length x 0.03 = Required DNA concentration in ng/µL).
  • Please supply at least 12 µL of template solution per reaction.


Primer Requirements

  • We supply some primers for use in sequencing reactions. The sequences of these primers are:

T3
5' -AAT-TAA-CCC-TCA-CTA-AAG- 3'

T7
5' -TAA-TAC-GAC-TCA-CTA-TAG-GG- 3'

SP6
5' -ATT-TAG-GTG-ACA-CTA-TAG- 3'

M13 Forward
5' -TGT-AAA-ACG-ACG-GCC-AGT- 3'

M13 Reverse
5' -CAG-GAA-ACA-GCT-ATG-AC- 3'

M13pUC Forward
5' -CCC-AGT-CAC-GAC-GTT-GTA-AAA-CG- 3'

M13pUC Reverse
5' -AGC-GGA-TAA-CAA-TTT-CAC-ACA-GG- 3'

  • The WDSF can also accept any complaint custom primer (as below). 
  • Please supply primers at a concentration of 5 pMol/µL (5 micro molar).
  • Please supply at least 2 µL of primer solution per intended reaction.


Sample Submission

Please refer to our Contact Details page for further advice on submitting your sample by post/courier.


Costs

For the latest equipment and analytical costs for external clients, please refer to the Science & Engineering Services Fees Schedule.

Tips & Troubleshooting

Discover our template tips, find solutions to common problems in template preparation and read our disclaimer.

Services

Check out the different kinds of services the WDSF has on offer.