WellGenetics is our preferred vendor for transgenic fly stocks.

Our primary contact is Dr. Yawen Chen (yawen.chen@wellgenetics.com). Say hi if you see her at the CSHL Neurobiology of Drosophila Conference!

Before ordering flies, make sure that BDSC and Janelia don’t have the flies made already. If the flies are potentially available at Janelia, you could ask Rachel to contact Barry Dickson lab for the flies and/or the plasmids that contain the enhancer fragment of interest.

If a transgenic service is needed and you have already obtained the plasmid for your fragment, you can use BestGene for injection services. Unlike WellGenetics that takes care of both cloning and injection, BestGene only performs injection and therefore requires you to ship them both:

1. The plasmid containing the enhancer fragment;
2. The vector from Addgene with Gal4, GDBD, p65AD or LexA, etc. inserted, depending on your need.

The whole process would be faster compared to WellGenetics.

Note: for generating plasmids to make your own transgenic flies see: /fly-stocks-and-crosses/plasmid-generation/

Starting a project#

Primer information#

For an enhancer fragment of interest, you will need to provide the appropriate sequences for the left and right primer.

• Primer sequences for the FlyLight/GMR collection are available from this  Excel file or csv file (courtesy of the FlyLight page of the Bloomington Drosophila Resource Center).
• Primer sequences for the Vienna Tiles collection are available as separate Excel files for current VT stocks and culled VT stocks (courtesy of the Vienna Drosophila Resource Center at IMP Vienna).

Plasmid type#

• WellGenetics already has the appropriate plasmid for making a plasmid for Gal4, GDBD, p65AD and LexA (pBPLexA::p65Uw).
• For other types of construct, you will need to obtain the plasmid from Addgene and ship it to them.

Landing site#

• You will need to specify a landing site for your construct.
• WellGenetics already has the appropriate fly stock for inserting transgenes into common sites, such as attP2 or attP40 site. For these sites, WellGenetics have a lot of experience, so they know how many embryos they need to inject to get at least one positive line. For less common site, it’s possible that they might have to do a few rounds to get a positive line (no extra cost, but longer calendar time).
• For other sites (e.g. JK22C), you may need to obtain the relevant landing site and ship it to them yourself.
• The main considerations when choosing a landing site are (1) ease of use in combination with other transgenic insertions of interest, and (2) the need to avoid transvection (Mellert & Truman 2012).

Balancing services#

• For an modest additional fee, WellGenetics will double-balance your transgenic construct for you.
• This is generally worthwhile as you need to remove things that are useful for screening anyway.

See this page on how to pay for the WellGenetics order (complicated!)

Intermediate checkpoints#

Once you place the order, WellGenetics will update you with the following reports. If you haven’t heard back from them, it might be a good idea to check the status of the project.

Cloning design report#

This part is fairly automatic (unless you want to specify a specific cloning strategy!).

Cloning report#

A month later, you’ll receive a report on whether the cloning was successful.

Note that the exact sequence of the enhancer fragment might deviate slightly from the reference genome. If there’s a larger deviation, WellGenetics will ask you whether it’s ok to proceed.

Sometimes, the initial cloning strategy fails, and they’ll need to change the cloning strategy (no additional cost, but extra calendar time).

Injection report#

You’ll get an injection report in 5-6 weeks after you get the cloning report.