Submitting Your Samples
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Completed sequencing reactions.
Reactions sets should be delivered to AgSci Bldg, rm 332 between 9am - 5pm
in a light tight container.
Please remember to denature your
samples prior to delivery by adding stop/dye solution, heating the
reactions to 92 C for 2 minutes and snap cooling on
ice. Longer denaturing
times should be avoided to prevent decoupling of the dye from the
fragments.
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Template DNA. Templates to be sequenced should be purified either
by organic extraction or through one of the various DNA purification columns
commercially available (we use QIAGEN products but others work equally well)
and submitted (minimum 50 ul total volume) at a concentration of 500ng/µl
in sdH20 or TE0.1 pH 8 buffer.
Several methods for estimating DNA concentration can be used for submitted
samples. These include: densitometry (spot density from ethidium gel), visual
estimation (from ethidium gel), UV quantification, and fluorimetry.
While fluorimetry is probably the most reliable, not all users have access
to this type of equipment and so we recommend densitometry (against known
DNA mass ladder standards). We do not recommend UV estimation because of
the effect UV-absorbing contaminants have on over estimating DNA concentration.
If users visually estimate DNA concentration from an ethidium gel, a photo
(which includes an appropriate moleculer standard) must be included
when submitting samples.
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E. coli colonies. Colonies should be streaked
and submitted on the appropriate selection plate such that single colonies
are visible on the plate. Mini preps (500 µl) can be submitted but an
additional 2-3 working days turn-around time should be expected while streaking
for single colonies is done in house.
* In all cases, indicate to lab personnel when and
where you drop off your sample and sample submissison form. Sample submission
forms are available in room 332 (or on our web page -
email sample submission form) and must be filled
out.
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