[
Worm Breeder's Gazette,
1980]
If you are peacefully picking worms one day and an eight-legged hairy monster walks into the field of your dissecting microscope, after you come down from the ceiling, you will know that you too have a mite infestation. You will also know the plot for a grade B horror flick. A piece of the moth crystals used in closets (paradichlorobenzene) placed in your incubator for a few weeks will kill them off with no detectable harm to worm stocks.
[
Science,
2002]
How much you eat, not what you eat, seems to make a difference in the aging process. It is well established that reduced calorie consumption robustly extends adult life expectancy in a variety of animal models. Now, on page 120 of this issue, Larsen and Clarke show that diet quality also affects aging. In the worm Caenorhabditis elegans, reduced consumption of coenzyme Q (Q) dramatically extends longevity.
[
WormBook,
2005]
Mutations in many genes can result in a similar phenotype. Finding a number of mutants with the same phenotype tells you little about how many genes you are dealing with, and how mutable those genes are until you can assign those mutations to genetic loci. The genetic assay for gene assignment is called the complementation test. The simplicity and robustness of this test makes it a fundamental genetic tool for gene assignment. However, there are occasional unexpected outcomes from this test that bear explanation. This chapter reviews the complementation test and its various outcomes, highlighting relatively rare but nonetheless interesting exceptions such as intragenic complementation and non-allelic non-complementation.
[
Worm Breeder's Gazette,
1989]
The map is now widely distributed electronically (see WBG 10(3), 67), but we are once again providing a summary for the gazette in the form of an output from the routine CHPLT. Do note that this is a provisional best guess, and that some linkages may later go away: please enquire if you need to know about the status of particular areas. When you receive cosmid clones, as stabs, please IMMEDIATELY streak them out on selective medium, pick small colonies, and grow 4ml minipreps (protocol from Alan Coulson if needed). For some cosmids, larger preps are liable to yield deleted DNA. Check that cosmid DNA appears full size (runs slower than lambda on agarose gels), then freeze a sample of good cells in 20% glycerol at -70 C. MRC computer account 'ARC' does not exist; Alan and John share account JES. A database node is now open at Seattle: modem number 206-467-2957; operator Phil Meneely. The summary of clone types given on the next page may be helpful when you are deciding which clones to request for your research. To reveal the most suitable clones for microinjection, the buried clones need to be displayed by the routine CONTASS; we will help you to do this if you ask. [See Figures 1- 3]
[
International Worm Meeting,
2007]
Imagine being able to run WormBase on your own computer, unencumbered from server load at www.wormbase.org, network traffic, or even the need for a network connection. Run BLAST searches on your laptop enroute to your next presentation or explore expression patterns of your favorite gene family at the corner coffee shop. Maybe you have a private dataset that you would like to incorporate with WormBase to share within your lab or organization. Or perhaps you would like to set up a mirror site of WormBase as a community resource. All of these scenarios are easily possible with WormBase packages called "virtual machines". Virtual machines contain the entirety of WormBase and an operating system to hold everything together. In essence, this WormBase-customized operating system runs safely inside your Mac (Intel), Windows, or Linux computer using free software from VMWare. Virtual machines are now created for every WormBase release. Learn more about how to use these on the WormBase Wiki:
http://www.wormbase.org/wiki/index.php/Virtual_Machines Bring your laptop by this poster and we will install the most recent version of WormBase for you!
[
Worm Breeder's Gazette,
1984]
In the last few months, several people have asked about the CGC strain freezing procedures. For general reference, here are some detailed notes and observations on freezing and thawing for optimum survival. Two important things. The most important thing is that you have lots of L1's and L2's to freeze, as they are the best survivors. L3's and L4's seem to survive at a lower rate, and adults do not survive well at all. The other important thing is that freshly starved worms seem to survive freezing better than well-fed ones. Getting lots of L1's and L2's to freeze. Homozygous stocks are generally pretty easy. If you start a large agar plate (15 x 90 mm, streaked with a grid of OP50) with ten L4 hermaphrodites, the plate will starve when the F2's have recently hatched. This is a very happy thing, and you can proceed to the actual freezing. With heterozygous stocks, you'll need to start a few clones on small (15 x 60 mm) plates, check the progeny for correct marker segregation, and pick three confirmed clones. When these starve, wash off all three plates and pool the worms for freezing. Other types of stocks are a little more tedious. These include burrowers, male stocks, paralyzed or severe Dpy and other slow-growers. With any of these types, it sometimes helps to start three or four small plates with one to five L4's, or three or four cross plates in the case of male stocks. As soon as you get a bunch of L1's and L2's on the plates, wash them off thoroughly, pool and wash the worms free of bacteria by centrifugation and removal of the supernatant. Resuspend the worms in 2 ml of M9 buffer and put them on a shaker at an appropriate temperature for 12 hours or so. Overnight is fine. This starves them. Before freezing, add one drop of an overnight OP50 culture so the worms will have plenty to eat when thawed, and freeze as usual. This isn't as much work as it sounds, and gives you a warm feeling later. It is especially useful with burrowers (most wild-type C. elegans isolates), of which there are usually none left on the surface when the plate starves. Actual freezing. Having obtained lots of larvae in a starved state, this is what I do: (1) Wash larvae off the plate with 2.2 ml of M9 buffer, draw the buffer/worm suspension into the pipette, measure ( usually 1.6 to 1.8 ml) and place the suspension in a sterile 30-40 ml test tube with closure; (2) add an equal volume of S medium plus glycerol (this is 30% glycerol, w/v, so the final is 15%); (3) vortex gently to mix; (4) draw up 3 ml in a sterile 5 ml pipette; and (5) dispense in 0.6 ml aliquots to five 2 ml Nunc freezer vials. These vials are placed in holes drilled into a styrofoam block which has been fitted into a styrofoam box that has a lid (Lew and Miwa, WBG 4: #1), and the whole thing is put in a -80 C freezer. Leave them there at least three hours; I usually leave them overnight. Then you can put them in liquid nitrogen. Four of the vials go into twin LN2 refrigerators; the fifth is put in liquid nitrogen in a separate place to be thawed the next day as a survival tester. Survival and contamination checking is done on the third day after thawing. If you have three small plates instead of one large, start with 2.5 ml of buffer to allow for what soaks into the agar. Wash one plate and draw the mixture back up, wash the next two plates with that. If you're doing the shaker trick, just add an equal volume of S + glycerol, vortex and dispense. It helps to use disposable graduated conical tubes with screw caps, since you can easily see how much S + glycerol to use. You can also prepare a single freezing solution, a 50/50 mixture of M9 buffer and S + glycerol, and use that to wash off the plates. You can either start washing with enough solution to leave 3 ml for dispensing into vials, or wash with a smaller volume and bring it up to 3 ml just before dispensing. Thawing. In my experience this is not terribly critical, but I try to do it the same way always. Vials are removed from liquid nitrogen and placed on the lab bench, and a timer is set for ten minutes. When the timer goes off, you'll usually have a chunk of ice with a layer of liquid around it in the vial. If it's still frozen solid, rub it briskly in your hands until the pellet starts to melt. Wipe the vial dry with a Kimwipe. This keeps condensed water from the outside of the vial from falling onto the petri plate. Vortex with the cap still on to mix the stuff into a runny slush and dump it into a clean, labeled agar plate. Results. At the beginning of the first Genetics Center contract, N2 freezing survival testers were frozen to provide monthly and yearly tests over the entire contract period. Survival rates from those testers averaged 26% with no decrease in survival in stocks frozen for the entire 5-year period; fluctuations around that average are presumably due to sampling variations. Long-term survival tests will continue. Survival of stocks stored at -80 C in a Revco is as good as in liquid nitrogen over an initial 6-month test period.
[
Worm Breeder's Gazette,
1993]
As some of you are already aware, Academic Press has asked Henry Epstein and myself to edit a volume of Methods in Cell Biology devoted to C. elegans. One of the useful appendices which we would like to include is a list of available antibodies and protocols for using them. If you have generated an antibody in your laboratory which you would like to see listed in a methods volume on C. elegans, please contact me as soon as possible. Incidentally, if you would like to make your antibodies widely available but are nervous about being swamped with requests, you might considered contributing your monoclonal antibodies to the Developmental Studies Hybridoma Bank sponsored by the National Institute of Child Health and Human Development. They are providing the wonderful service of growing up monoclonals and making the cells or supernatants available to researchers at reduced fees. I know from personal experience that their anti-tubulin antibody works well for C. elegans, and it is considerably cheaper than commercial products. Contact Dr. Thomas August, Johns Hopkins University at (410) 955-3985 for more information about receiving a catalog or making contributions. I have the impression that they are not anxious to handle hundreds of worm specific antibodies, but that they are interested in handling a limited set of the most widely used C. elegans specific antibodies as well as antibodies which are known to cross-react with other species.
[
Worm Breeder's Gazette,
1995]
As many of you know, there is a gopher server at the CGC(elegans.cbs.umn.edu) and a World-Wide-Web (WWW) server in my lab inDallas
(http://eatworms.swmed.edu/) devoted to information about Celegans. These servers are heavily used.There are several other C elegans information servers, but they are notincluded here because I don't have their usage information.The table shows that, although many connections come from net-surfers whovisit once and never again (perhaps having expired from boredom), there isa large group of repeat offenders who use the servers over and over. Many, perhaps most, come from C elegans labs. (For instance,wormworld.ucsf.edu and horvitzlab3.mit.edu are pretty obvious.)My point is, if you need to reach the C elegans community, this is a goodway to do it. There is an Announcements page on both gopher and WWW. Inthe 55-day period analyzed it was read 203 times. If you have a jobopening or a conference or anything else you'd like worm people to knowabout, you probably should be advertising there. Announcements can besubmitted by e-mail to leon@eatworms.swmed.edu, and usually appear withina few hours.