1. Make sure that you have a USB key with the e-gel images saved on it.
  2. If you forgot to brighten the image when it was on the computer in the darkroom, just open it and bring the brightness (small picture of a sun) up to about 90%, or until it looks bright enough (you can also brighten the image in E-Editor).
  3. Open E-Editor. Go to “Open” on the top left corner, and find the picture that you wanted to put on LIMS.
  4. Go to the left hand side of the screen once your image is there, and press on the “Crosshairs” left hand side. This will give you three red crosshairs. Line those up with the white crosshairs on your e-gel.
  5. Once the cross hairs are aligned, hit the “Frame” button on the left hand side, and green lines will show up on the page. Click on the green lines and drag them until they are arranged so that the white bands on your e-gel are within the green lines.
  6. When everything seems to line up, hit “Next” at the top of the screen. It will rotate your image a slight amount. If the image still looks ok, click “Next” again. If it looks misaligned, try moving the frame again.
  7. You should end up with an image that is 8 by 13 wells. Save it as a jpeg file.
  8. Go to LIMS, under the section for e-gel images PCR Gel Check chose Gel Image Upload.
  9. User Barcode: Scan in your card. Plate barcode: scan the PCR plate. Protocol: select E-gel (it is the only option).
  10. Upload the image of the e-gel.
  11. You have to look through each image and select:
  • Band number – select 1 if there is a band (you want to look for the band closest to the left side) or 0 if there is no band. Rarely do you ever select 2 since that is for a double band.
  • Intensity – Rate the intensity from 0 (no band) to 4 (very bright band).
12. Select the reuse count – that is the number of times the gel was reused – therefore if the gel is brand new, the reuse count is 0.

Return to E-gel image on the Computer.

The next step is either Sequencing Procedure For BIO or Sequencing at the Genomics Facility depending on which one you are supposed to do.

Updated April 27 2009