THE FOLLOWING REQUIREMENTS MUST BE MET BEFORE YOUR SAMPLES CAN BE SUBMITTED

PLEASE NOTE: Due to continuing concerns posed by potential bioterrorism agents (select agents), our online submission form requires that you provide us with information on the original biological source of any DNA from which you will be asking us to obtain sequence. "Original biological source" refers to the organism from which the DNA was first found in nature (human, HIV, Staphylococcus aureus, etc.). If you simply put a nonspecific term such as "E. coli clone", this will not be sufficient. We realize that adding yet another field to fill out on these forms can be tiresome, but this information is necessary to protect our staff against potential legal difficulties. Thank you very much for your understanding.

            - David W. Dyer, Ph.D.

 

Preparation of samples:

1. All samples must be submitted in 96-well format (V-bottom plates, skirted or non-skirted PCR plate, or Optical Fast plate).
2. If you are submitting <96 samples, then load the plate in column format (i.e. A1-H1, A2-H2 etc.) Partial plates can be reused for submission, in which case you can use the first available column (i.e. starting with A4-H4 etc.) Just be sure that if you start with A4 on your plate you also begin with A4 on the submission form.
3. Follow Applied Biosystems instructions according to what type of reaction you are doing. Each well should contain at least DNA and primer. If you are having us add the Master Mix please select that option on the submission form.

4. An SUR# (OUHSC) or PO# (off-campus) must be given at the time of submission. There is a tray in the main office where you drop off your submissions that is labeled "SUR." We bill every month on the 15th, so you will need to submit a new SUR or PO every month if you are submitting samples. Without a current SUR or PO, your samples will not be run.

   For samples that need to be shipped, if there is no Master Mix already added, please dry down your template and primer mix in a PCR plate before sending to reduce the chances of sample loss during shipping. If there is Master Mix already added, ship the samples cold (4º C).

5. An online submission form (initially, create a new account and choose the appropriate service, otherwise sign in and procceed) must be filled out completely.
6. We recommend that you have set up reactions in duplicate and also always run a NTC (No Template Control) for each primer pair.
7. If you are running TaqMan reactions, bring down and drop off or email Mandy Gipson the taqman files that you received from Applied Biosystems.

Shipping Address:

The Laboratory for Genomics and Bioinformatics OUHSC
Attn: Mandy Gipson
975 NE 10th Street, BRC1106
Oklahoma City, OK 73104

TO KNOW:

1. Turnaround time for customer reactions is dependent on the number of samples we receive each day. Customers can expect their data within a maximum of 48 hours from the end of the day that the samples are received (not including weekends, or holidays). For example, samples that are received on Monday by 4pm will be ready for download no later than 5pm on Wednesday. Larger orders of multiple plates may take longer than 48 hours to complete, but data will be made available for download as each individual plate is processed.
2. Finished data will be emailed to the address on the account unless specified otherwise.
3. If you submit plates containing gaps between samples within a column you will be charged the regular reaction rate for these gaps.
4. Once the samples have been run, plates with empty wells will be stored on top of the refrigerator where submissions are placed for pickup. If plates are not picked up within two weeks, they will be discarded. We ask that people only take back plates that were submitted by them. We cannot be held responsible for lost or stolen plates. If you have any questions, please feel free to contact Mandy Gipson.
5. All reactions will be cycled under the typical thermal cycling conditions recommended by Applied Biosystems unless you have noted otherwise in the comments section.
   If your assay uses TaqMan Gene Expression Assays or Custom TaqMan Gene Expression Assays probes and primer and the TaqMan Univeral PCR Master Mix, No AmpErase UNG Kit:

   Initial steps	PCR (Each of 40 cycles)
   AmpliTaq Bold DNA Polymerase Activation	Melt	Anneal/Extend
   Hold	Cycle
   10 min. @ 95oC	15 sec. @ 92oC	1 min. @ 60oC

   
   If your assay uses probes and primers designed with Primer Express software and uses the TaqMan Universal PCR Master Mix Kit:

   Initial steps		PCR (Each of 40 cycles)
   AmpErase UNG Activation	AmpliTaq Gold DNA Polymerase Activation	Melt	Anneal/Extend
   Hold		Cycle
   2 min. @ 50oC	10 min. @ 95oC	15 sec. @ 95oC	1 min. @ 60oC

   
   
6. The recommended reaction volume ranges for Applied Biosystems Instruments are as follows:

   Instrument	Plate Type	Minimum Reaction Volume (ul)	Maximum Reaction Volume (ul)
   7500 Standard	96 well (.2mL)	25	100
   7500 Fast	96 well (.1mL/Fast)	10	30