- Create an account with the DNA LIMS of the Ohio University
Genomics Facility. You can do this by clicking on the Request and Reservations link in
the home page.
- Remember the completed sequencing results will be mailed
back to the email address provided during your account creation.
- Follow the instructions in the sequence submission page.
- Quantify your DNA samples before you submit to the facility
- Recommended DNA concentrations:
| PCR Product |
Quantity |
| 100-200bp |
1-3ng |
| 200-500bp |
3-10ng |
| 500-1000bp |
5-20ng |
| 1000-2000bp |
10-40ng |
| >2000bp |
20-50ng |
All PCR products need to be cleaned prior to submission.
Before submitting a plasmid DNA the concentration is
dependent on size of the plasmid DNA + insert. The general principle is
25ng/ µ l per 1000bp (for example if you have a plasmid of
3000bp and insert of 1000bp your total plasmid size is 4000bp, so you
need 4 X 25ng/microl = 100ng/ µ l).
- Please submit at least 5 µ l of your DNA sample.
If you want to reclaim the remaining template you can request the
facility to save it by mentioning that in the comments section of your
DNA sequence request form and collect it from the facility after the
sequencing is done.
- Do not forget to put the request number on the DNA sample
tube. The request number is unique and generated when you submit your
sequence request form. If the same template is used for multiple
reactions you can mention either the range or all the request numbers
on the tube.
- The facility provides the universal primers like M13
forward, reverse, T7 promoter, T3, SP6 primers for no extra cost.
- If submitting your gene specific primers, please submit at
least 5 µl of the primer per reaction at the concentration
of 3.2 pmoles/ µ l. Please indicate on t
he tube the primer name and the request number/numbers that the primer
should be used for.
- Please mention in the comments section of the sequencing
request form if any of your DNA samples are GC or GT rich as the
facility would process those samples using a different chemistry.
- Please bring the DNA samples and primers (if you are
sequencing using gene specific primers) along with a copy of the
sequence request form to 510 PORTER HALL. You can leave the samples and
primers in the -20C freezer situated at the back of the lab and leave
sequence request form in the bin above the freezer.
- Please give the facility about 1-2 days to process your
samples.
Microarray
- Contact Vijay Nadella to discuss about Microarray services (email:nadella@ohio.edu)
How to use Quantitative PCR machine
- Create an account with the DNA LIMS by clicking on the Login link.
- Use Core Calendar to access the Stratagene Mx3000p qPCR machine usage.
- Check the access request box so that the administrator can grant you an access to schedule the usage
- The restriction will allow the administrator to properly train the individual before they can start using the machine
- If you will be running optimization run please indicate that in
the comments section of your reservation as the optimization run will
not be charged
- Please fill in an appropriate account number when you schedule the usage of the q-PCR machine
How to submit Bioanalyzer samples
- If you are first time user of the Bioanalyzer services
please check with the Core staff regarding the availability of Chip of
your interest
- Create an account with the DNA LIMS by clicking on the Login link.
- Click on the Bioanalyzer tab and fill out the appropriate chip sample reqeust.
- Provide atleast 3-5 ul of your sample in 0.5ml tubes.
- Sample tubes need to be labelled with the requistion numbers and the sample names (if possible)
- Please bring your samples along with a printed sheet of the Bioanalyzer sample sheet to 510 Porter hall
- Please leave your sample to be processed by Bioanalyzer in the
third shelf of the freezer in the Genomics facility and leave the
sample sheet in the document bin on top of the freezer
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