SREL
DNA Lab
Option 1 - Enrichment for Microsatellites
Overview:
E-mail Glenn@srel.edu to determine when/if we can
schedule you in.
Send us:
2 tubes of DNA per
species (≥100 無 of >50ng/無 of high molecular weight DNA)
a signed work order
Here are the details:
1) Download a copy of our enrichment protocol from the SREL DNA Lab Protocols web page. You are free to
follow the protocol yourself, or we can do the enrichments for you; you will
want to have the protocol either way.
2) You will have to do step I - Isolating the DNA. You will need to
isolate ~10 ug of high molecular weight DNA (ideally more than 100 無 of
>100 ng/無, but 50 ng/無 will work fine; and even 20 ng/無 usually works).
We recommend sending 2 or 3 good DNA samples per species of interest. You can
use Qiagen or Wizard or PureGene kits or do PCI extractions - do whatever works
best for you. Before sending the DNA to us, run out an aliquot of DNA on a 1%
agarose gel along with lambda DNA - about 90% of the DNA should be the same
size as the lambda DNA. If you
have questions, e-mail Travis (Glenn@srel.edu).
3) We will do steps II - V: Digesting the DNA, Linker Ligation, Enrichment
with Biotinylated Probes, and Recovery PCR. Please see the current list of probes
available. We do serial/double enrichments with 3 different probe mixes.
A variety of probe
mixes are available, but we will generally only modify the mixes used
for large-scale projects (i.e., projects needing several hundred primer pairs).
4) We will send you:
a) DNA eluted after enrichment,
b) PCR amplicons from the eluted DNA (i.e., recovered DNA),
c) SuperSNX-f so you can recreate (b) from (a), fresh for TA cloning, and
d) copies of all gel pictures.
5) We now guarantee that at least one of
the three libraries will yield >25% of clones with microsatellite repeats.
Most projects will yield at least 2 libraries with >50% of clones with
microsatellite repeats. Please
note, however, that many genomes contain highly repetitive elements (e.g.,
transposons) with microsatellites and these can represent large numbers of
positive clones in any one library.
This is why we do separate enrichments with moderate numbers of
oligonucleotides in each.
6) Please refer to the Order Form for
current prices. These are considerably less than commercial sources. As part of
this "bargain" you may have to tolerate potential delays on our part
(we are generally doing enrichments about twice a month), but sometimes we are
traveling, etc. If you enquire
about potential schedules prior to sending your DNA, we will do our best to stick
to the target schedule agreed upon. Gentle e-mails requesting updates every few weeks are welcome
and often help move things along.
Website content by Travis Glenn
For more information, contact: Glenn@srel.edu
Back to Microsatellites - home.
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Option 1 - |
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Launched 14 December 2001
Revised 2 April 2007