I have plasmids that I want to send for sequencing, but I want
to make sure that I have the right concentrations for the plasmids
and the primers:
– The concentration of the plasmid 1(7000bp) = 109 ng/ul,
forward primer (657 bp) = 9.8 ng/ul, and reverse primer (410 bp) =
30.2 ng/ul
– The concentration of plasmid 2( 7000 bp) = 84.2 ng/ul, forward
primer(658 bp)= 20.3 ng/ul, and reverse primer( 408 bp)= 15.7
ng/ul
* The previous concentrations should be prepared according to
the table below:
Note:
The plasmids will be sent with only one primer ( either
the forward or the reverse primer), and the complementary primer
will be added by the company.
For example:
If I send plasmid 1 with the primer in the forward
direction, the company adds the primer in the reverse
direction.
US &EU PreMixed In the same tube, mix template (10㎛ and your primer (5 according to the table below. To use a GENEWIZ Universal Primer, simply submit template at the requested concentration in 10 See the Technical Notes section for tips on how to purify PCR Premixed Volume” Template Total Mass Your Primer Total Picomoles DNA Length (include vector) DNA Type Template Concentration in 10 Template +Your Primer) -500 ng 800 ng -1000 ng -10 ng 20 ng 40 ng 60 ng Treat as plasmid -50 ng/ -80 ng / μ -100 ng /pl ~1 ng/μ 25 pmol 15μ1 6-10 kb Plasmids 10 kb 500 bp 500 1000 bp2 ng /l 1000-2000 bp 4 ng /pl 2000 4000 bp-6 ng /pl 4000 bp 25 pmol 15μ1 Purified PCR Products Treat as plasmid
