Template FAQ

How much plasmid DNA do you need for a standard premixed tube reaction?
We require 600 ng of template for each sequencing reaction. This amount is mixed with 2 µl of 4 µM primer and brought up to 18 µl total volume with H2O.

How much plasmid DNA do you need for a high volume premixed 96-well reaction?
We require 600 ng of template for each sequencing reaction. This amount is mixed with 2 µl of 4 µM primer and brought up to 18 µl total volume with H2O.

How much PCR product do you need for a standard premixed tube reaction?
We require10-20 ng of PCR product for every 200 bp of product per reaction. This amount is mixed with 2 µl of 4 µM primer and brought up to 18 µl total volume with H2O.

How much PCR product do you need for a high volume premixed 96-well reaction?
We require 5-10 ng of PCR product for every 200 bp of product per reaction. This amount is mixed with 2 µl of 4 µM primer and brought up to 18 µl total volume with H2O.

Should I resuspend template in TE or H2O?
Either is fine. H2O is perhaps better.

How long is the average read?
Template quality will affect length of read. Average read length is 500 bp, however, lengths of  650-750 bp are often achieved.