Single Read Sequencing - Sample Requirements
Concentration:
Plasmids: 30-100ng/µl
Cosmide/BAC/PAC: >100ng/µl
PCR fragments: 10-50ng/µl
Custom primers: 10pmol/µl
Volume:
DNA samples & primers: 20µl each (sufficient for 8 reactions), for 96 well sequencing please send 50µl in a single tube. Please send the DNA dissolved in water. The solution must not contain any EDTA. DNA and primers to be delivered in 1.5ml reaction tubes or 96 well microtitre plates.
Primer conditions:
The melting temperature (TM) of the primer should be between 52°and 58°C and the length should be between 17-19bp. Ideally, the GC content of an 17mer should be 7-9 G+C; for an 18mer 8-9G+C and for a 19mer 10 G+C.
G or C should be at the 3’ end, but not more than 3 Gs or Cs. The primer sequence should be a good mix of all 4 nucleotides with no more than 4 identical bases in a row (AAAA or GGGG).
Storage of DNA and primer:
Storage of DNA and primers delivered by customer: 10 days
Storage of custom primers synthesized by GATC Biotech: 1 year
