Last updated: 01-05-2011
Sequencing Protocol
Remember:
If you want sequencing to be picked up that day, your samples MUST be in the dropoff box on the first floor of 68 by 3PM. The box is located near the DHL and FedEx mailboxes near the main stairs.
Samples must be prepared in 8 strip PCR tubes. If you’re using less than 8, cut off the empty tubes for future use.
Preparing your samples:
In the sample tube, mix 10 ul of your template (at specified concentration - see below for plasmids vs. pcr products) and 5ul of your primer.
Plasmids
Length |
Template |
Primer |
Total |
|---|---|---|---|
<6 Kb |
~ 50 ng/ul |
5 uM |
15 ul |
6 - 10 Kb |
~ 80 ng/ul |
5 uM |
15 ul |
>10 Kb |
~ 100 ng/ul |
5 uM |
15 ul |
PCR products
Length |
Template |
Primer |
Total |
|---|---|---|---|
<500 bp |
~ 1 ng/ul |
5 uM |
15 ul |
500 - 1000bp |
~ 2ng/ul |
5 uM |
15 ul |
1000 - 2000 bp |
~ 4 ng/ul |
5 uM |
15 ul |
2000 - 4000 bp |
~ 6 ng/ul |
5 uM |
15 ul |
> 4000 bp |
treat as plasmid |
treat as plasmid |
treat as plasmid |
Genewiz has a list of universal primers that your vector may have and they will add these for you at no added cost. If you choose to use these, only add 10ul of your template.
Before submitting, you must place your sequencing order online. The online ordering system will create tube labels using your initials and the sample number. Use these labels on your tubes.
Print a copy of the order receipt and send it along with your samples in an sealed and labeled envelope.