McClean:Sequencing

We use Macrogen for sequencing plasmids and DNA fragments. It is very convenient, as the sample can be dropped off in the fridge in LTL (next to where we pick up primers) and the sequencing is generally done by the next day.

To sequence DNA with Macrogen:

• Go to http://orders.macrogenusa.com
• Set up an account
• Request a PO for your sequencing order using the Princeton https://genpurch.princeton.edu system (write in the notes section that you will send in the PO yourself, figure about $5 per reaction)
• Prepare your samples (see http://orders.macrogenusa.com/support/seq/sample_pre.jsp for details on how to prepare your sample):
  • We generally use the 8-strip PCR tubes (available in the stockroom)
  • Our oligos are at a concentration of 10μM, so to get the desired 8pmol of oligo, dilute our oligos 1:5 in sterile water (for a final of 2pmol/ul) and add 4μL to the 8μL of DNA to be sequenced.
• Once you receive your PO, complete an on-line Macrogen order form and use the PO number as the payment option. Use:

Princeton University Purchasing Department P.O. Box 33 Princeton NJ 08543-0033 as the billing address.

• Place your samples together with a copy of the order form in the drop off refrigerator in front of LTL-136. (You can use any small, clean, ziploc bag to submit your sample in, check the box-o-bags above the Plasma Preen)
• Retrieve your data from the Macrogen server the next day. You will get an email when your sequencing is done.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

1. List troubleshooting tips here.
2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
3. Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.

*Megan N McClean 19:24, 8 September 2012 (EDT): This protocol was changed on 9/8/2012 based on email from the SYnseq facility at Princeton notifying use that we no longer needed to request a PO, but could instead use our account number. See McClean:Orders_from_IDT/Genewiz/Macrogen/GenScript for more information. The old protocol is in the comments (visible in edit mode). Don't get rid of it if you edit this file please.