DNA Sequencing - Sample Prep

(Effective Date: Monday, October 14, 2002)

To submit samples, you can either premix the template and the primer, or send them separately. However, please do not premix for the first a few times, so that we can check your template and calibrate the concentrations (free for the first 3 times for new customers). The followings are the conditions for both premixing and non-premixing sample preparation. When possible, please use 8-strip PCR tubes for <96 samples and primers, and use 96-well plates for > 96 samples:

I. If you are not premixing the samples, please send according to the following specifications:

1. Non-premix Template (in water):

	PCR products: concentration >10ng/ul, please send >10ul and let us know the concentration and the length of the PCR product.
	Plasmids: concentration >50ng/ul (100ng/ul is preferred), please send > 10 ul each.

2. Non-premix Primer (in water): concentration 3 uM, please send > 10 ul each, and have enough for all the samples.

II. If you premix the template with the primer:

1. Premix Template (in water):

	PCR products 100-200bp     2-3 ng 200-500bp     6-8 ng 500-1000bp   10-15 ng 1000-2000bp  20-30 ng >2000bp        60-100 ng
	Single-stranded: 100-150ng
	Double-stranded: (plasmids) 500ng

2. Premix Primer (in water): 8 pmole
3. Premix Total volume (in water): 15 ul.

For Non-premix samples, we have the following additional services available. We can prepare templates at $10/each (miniprep for plasmids, or purification of PCR products). If you are not sure about the template concentration, we can check it for you at $2/each. For templates with low, but reasonable concentration, we may also concentrate the sample before the run at $5/each.

We have M13 forward (-20); M13 forward (-40); M13 reverse; T7, T7 reverse, T3; and SP6 available as common primers. However, for the less popular common primers (M13 reverse -40; Lambda GT11 forward and reverse; SK; KS; 5' or 3' pKK; pET; etc.), if you anticipate that you'll use them a lot and would like us to stock them for your non-premixed samples, we can do that after confirming the primer sequences with you.