Note: for having transgenic flies made see:

  • /fly-stocks-and-crosses/wellgenetics-services/
  • or, consider making a BestGene page to document that service (preferred by some).

Genscript#

Genscript will synthesize and clone plasmids for generating flies. Compared with Twist Bioscience, they are somewhat more expensive and slower, but they will synthesize sequences that Twist will not. They are probably your best bet if you’re trying to synthesize enhancer elements (to make driver lines). Genscript also does custom cloning if you already have your DNA of interest and just want to move it to another vector.

Login#

Username: flypokers@gmail.com

Password: wilsonlab1

Twist Bioscience#

We have most recently used Twist Bioscience for making plasmids to generate flies. For our purposes, Twist only does synthesis of a DNA fragment into a target vector. That is, if you already have DNA in hand containing your sequence of interest, you only need cloning services, and Twist will not do that. Use Genscript instead. Twist has pretty strict requirements for what sequences they will synthesize, though when they do make them, they are cheaper and probably faster than anyone else. Twist is probably best suited for synthesizing effectors. That is, protein-coding genes are better behaved from a synthesis perspective and codon-optimization allows some flexibility in the exact DNA sequence. For synthesizing enhancer elements, try Twist first, but don’t be surprised if the system complains about repeats, CG content, etc. and refuses to make it.

Login#

Username: flypokers@gmail.com

Password: wilsonlab1!

Services#

Twist’s Clonal Genes service involves synthesizing the DNA sequence of interest into a target vector. Under our custom vectors, we have commonly used ones for transgenic flies:

  • pJFRC7-20xUAS-IVS - for making 20XUAS-your synthesized effector. Plasmid has attB site for landing into attP site in flies. Thank you to the Clandinin lab for sharing this vector with us in the Twist system.
  • pJFRC19-13xLexAop2-IVS - for making 13XLexAop-your synthesized effector. Plasmid has attB site for landing into attP site in flies. Thank you to the Clandinin lab for sharing this vector with us in the Twist system.
  • pBPLexA–p65Uw - for making your synthesized enhancer-lexA::p65AD (i.e. for making a lexA driver line)
  • pBPGAL4-2Uw-2 - for making your synthesized enhancer-GAL4 (i.e. for making a GAL4 driver line). Vector onboarding in progress (11/1/23)
  • pBPp65ADZpUw - for making your synthesized enhancer-p65AD (i.e. for making an AD split-half driver line). Vector onboarding in progress (11/1/23)
  • pBPZpGAL4DBDUw - for making your synthesized enhancer-GAL4DBD (i.e. for making a GAL4DBD split-half driver line). Vector onboarding in progress (11/1/23)

Purchasing#

To make a purchase you can pay by credit card or PO. Note this can get expensive, so make sure the P/Pro-card isn’t close to its limit when attempting a credit card purchase. If it is a PO, you have the option to generate a quote with frozen line items that can be uploaded to B2P (HMS) or P2P (HHMI). If you choose PO, there are a couple ways to get the PO to Twist. Once the order is approved you can ask to be sent (or generate yourself if you’re a purchaser) a PO to email to twist yourself, the checkout page outlines these nicely.

If using a PO, make sure the billing address is the following:

Accounts Payable

Howard Hughes Medical Institute (HHMI)

4000 Jones Bridge Rd,

Chevy Chase, MD 20815

email: flypokers@gmail.com

phone: 6174325571

Also, ensure that HHMI is somewhere in the name on the quote, e.g.:

Rachel Wilson

HHMI / Harvard Medical School

200 Longwood Avenue

Boston, MA, 02115