Endy: DNA sequencing
How to submit a DNA sample for sequencing:
- Go to the Biopolymer lab website and read the page on DNA sequencing.
- Prepare your samples:
- Samples should be prepared in 8-strip PCR tubes with lids.
- Samples should not exceed a volume of 12µl; calculate ahead of time how much water, DNA, and primer you will add.
- First, add the water. If using a custom primer (like VF2 or VR), add your primer (final amount should be 3.2pmol; use 2µl of a 1.6pmol/µl dilution of primer). Finally, add your DNA (see Biopolymer lab website for amounts; use 200-300ng of plasmid DNA).
- Once everything has been added, close the tubes w/ a strip of caps.
- Go to dnalims.mit.edu and login.
- Go to Enter DNA sequencing requests.
- Input the number of samples and fill out the rest of the sample information. Use the Endylab account # for the order. Be careful not to mix up your samples. You want to be sure you know what sequences that you get back go with which samples.
- Submit the request and print two copies of the receipt. Keep one for Endylab records and take one with you to the sequencing facility.
- Bring your samples and printout to the sequencing facility (4th floor in the cancer center; enter by the entrance opposite the south end of bldg 68). Put your samples in one of the empty falcon tubes, label the tube w/ the order # and your name, and leave the tube in the fridge. Put your printout in the sleeve on the fridge.
- When you get the email saying your sequences have been completed, log into dnalims.mit.edu and go to retrieve sequences. Download the chromatogram and analyze it using ApE.