Volume reaction

For each reaction, we need 5 uL, as the following composed:

n μL Template DNA
0.5 μL Primer at 10 uM
4,5 - n μL H2O

Template DNA

Depending on the type of template, please supply the indicated amount of material per reaction DNA (5 μL)

Plasmid DNA < 9 kb 300ng
Plasmid DNA 9-10kb 500ng
Plasmid DNA >10kb 500ng*
PCR product 2ng per 100bp

As the DNA template is one of the main factors influencing sequence quality, it should be purified using commercial kits and diluted in distilled water (or Tris), free of EDTA and other salts, proteins, RNA, or genomic DNA.

PCR products have to be purified from reaction buffer, primers and nucleotides. They must appear as single band in an agarose gel. PCR purification is available at our facilities (please contact the Sanger team for more information).

Primer

0.5 uL of specific primer (10 uM) per reaction.

Primers should be added into the mix by the customer .

* Please note that in this case we do not guaranty the sequencing results. Therefore we highly recommend using our modified protocol or Full service. For more information please contact us.

Fasteris | DNA Sanger One Shot Services