How tos and FAQs

CleanSeq – Preparation of samples

Sample is an unpurified PCR product (aliquot of the PCR reaction) or other suitable template, amenable to enzymatic cleanup. You must also supply sequencing primer(s) or select some of our universal primers.

 

Primer requirements:

  • Primer volume: 200 μl per 100 samples.
  • Primer concentration: 5 μM.
  • Please note that up to twelve sequencing primers can be used if samples are supplied in the plate. If samples are supplied in tubes, one primer can be used for each eight samples ordered.

 

Sample requirements:

Template

Concentration

Total volume

PCR product <500 bp

~10 ng/μl

5-10 μl

PCR product 500–1000 bp

~20 ng/μl

PCR product >1000 bp

~40 ng/μl

Plasmid

~100 ng/μl

 

Note: If you want to order bidirectional sequencing, it is just enough to send for example a single plate with 96 samples, each having a volume of 5-10 μl, and two sequencing primers. But it is necessary to order sequencing of two plates - the first plate by the CleanSeq protocol (using one of primers) and the other by the StandardSeq protocol (using the other primer).

We recommend 8-tube strips with individual caps (eg Eppendorf 0030124359) or 96-well plates. Do not use parafilm. Please mark the tubes preferably with a number indicating their order in the order form.

Download sample prep guide in pdf.

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