PCR Purification Submission Policy

PCR purification is a 96 or 384 well plate format service, which is available for researchers who are experienced users of the facility and are confident in their methodology for PCR and primer preparation. We sequence and charge for all 96 or 384 wells on the plate regardless of the results, unless it is a clear technical error on our part. Acceptable plates are listed below:

  1. 96-well format: Eppendorf Twin Tec semi-skirted plates, part #951020389, they are located in the Yale stockrooms, or can be ordered directly from Eppendorf. Also, Applied Biosystems Optical 96-Well Reaction Plate, part #N801-0560, these are also available in the Yale Stockroom

    OR

    384-well format: BioRad Hard-Shell 384-well Skirted PCR Plates, part #HSP-3801. Also, Applied Biosystems MicroAmp Optical 384-well reaction plates, part #4309849.
  2. PCR purification plates should be submitted with a printed PCR Purification online submission sheet
    (Yale, Non-Yale)
  3. After PCR Purification, the plate is sequenced by our standard sequencing protocol.

PCR Purification Plate Submission Protocol

Add to each well:

20 µl containing 15-25 ng/µl of UNPURIFIED PCR product

For Primer Submission:

  1. Tube format: 384ul of 4µM custom or universal primer in an eppendorf tube for plates that will use just one primer for entire plate, or:
  2. 96-well format: 5 µl of 4µM custom or universal primer/well. Primers should be oriented according to PCR plate layout.

Cap the plate firmly using the strip caps/or sealing film, wrap each plate with parafilm.

Be sure to clearly label each plate as either PCR Purification plate or Primer plate with a unique name. The Primer plate should reference the corresponding PCR Plate for sequencing.

Once we have purified your PCR product, we will then add your primer and sequence your PCR product. Results will be posted in your folder via FTP.

Results will be labeled with the researcher code - plate name -and well-coordinate (RES CODE-plate name-A1-12, RES CODE-plate name-B1-12 etc). The researcher is responsible for holding the "key" to the well co-ordinates for sample identification.

Standard Turn Around Time

Our turn around time for a single plate is 24-36 hours from the time we receive your plate (except for plates received on Friday). Please allow extra time when you submit more than one plate at a time.

For any questions or concerns, please contact the lab directly, 203-737-2566 or dnasequencing@yale.edu