How to start a "grant club"

Starting a lab is sometimes a lonely pursuit. You are now in charge of everything and must have the final say on papers and grants that go out. While it may have been like this even before your transition, now your grants do not benefit from the shelter of your mentor's name and the success of your new lab depends on them.

Finding good readers becomes really important, because you need people who may not fully understand your field to take a serious look to your proposal and give you feedback. The more detailed and honest it is, the better. While senior investigators with experience in obtaining NIH funding will help, sometimes they have time only for a passing look and you are still wondering whether there is something you should change. Since there are eight of us who were hired in the past few years, we organized and started a monthly "grant club", where someone submits their latest effort for slaughter. R01 aims or full proposals, foundation grants or career award applications are sent out to everyone and dissected over pizza and beers. After 6 months everyone seems to really like it, in particular the fact that we all provide a lovingly brutal sounding board to test ideas. Now we started adopting stragglers from other departments, who have an interest in neuroscience.

The recipe is simple, 1) make a Doodle and find a day when most people can come, 2) the host/grant writer sends out their draft a few days before, 3) the host gets pizza/beer or fancier fare and other people can chip in (Girls, if I guy is hosting, remember the fruit and cookies...), 4) to break the ice you can ask for advice on how to get students or can just start complaining about things you would like to change in your institution, 5) discuss the grant. Do the aims weave a good story? Is the questions asked approaching a fundamental mechanism? Can you even tell what the fundamental mechanism studied is?   Is it too much work? Too little work? Is it cohesive? If Aim 1 doesn't work, will everything fall apart? and so forth and so on. Be nice and honest, but more importantly try and be helpful.

Which brings me to a sticky issue, be mindful of the purpose of these meetings to find the right mix of colleagues. As far as I can tell, we all really care about each other's success and we all enjoy discussing each other's science. Many of us don't have a chance to talk apart from this monthly occasion and it is great to bring everyone together. You may already have a wonderful group of colleagues that have offices right next to yours and that you meet often for coffee. Or you are in a small place where there are only a couple new hires or in a big place where everyone is competing and spying on everybody else. Then I would say, propose an online grant club with friends at other universities or meet with people at other local institutions. It makes such an enormous difference to have a cohort to lean on and if you have no grants coming up, meet anyways to talk about science, lab management or
which grants people have applied to.

Biotium GelRed and GelGreen to replace ethidium bromide

We would like to run agarose gels without ethidium bromide to avoid the need of dealing with toxic waste and to reduce the use of dangerous chemicals in the lab. We talked to quite a few vendors about this and started trying things out.

We started with AMRESCO EZ-Vision dyes which are SYBR Green based and were not very happy with them. Staining was very weak and we could not reliably image the gels on our existing UV image acquisition system without a SYBR filter. We had no intention of buying a new image acquisition system just for this so we kept looking. 500uL $89.52 from VWR

We tried Promega Diamond Nucleic Acid dye was also pretty weak and it is not an in-gel stain, so requires additional time for post-staining. We decided to keep looking. 500ul $70.00 from Promega

Finally, we started having good results with Biotium GelRed. GelRed is very sensitive and looks orange/red on the UV transilluminator like EtBr. GelGreen was not quite as bright, so we focused on GelRed, but optimization was needed. We had two primary issues: 1) bands which are very close to each other appear like they are fused together and the molecular weight marker was just a smear, 2) bands sometimes ran at a different weight than expected.
Since one of our genotyping PCRs needs to distinguish between a 400 and 480bp band it was very important to get good separation. Biotium recommended to reduce the amount of dye in the gel or to try post-staining because both the separation issue and the run length issue could be due to the larger size of the Biotium dye which can impair DNA motility. We lowered the amount of in-gel stain from 10,000X to 40,000X and it still worked pretty well, but a range of 20-30,000X may work best (around 2ul for a 50mL gel). Another way to improve separation is to post-stain if necessary and post-staining is relatively quick (15-20 mins) and does not require destaining. In addition the post-staining dye can be reused saving some extra money. Because of the size of the dye, agarose concentration is also critical and concentrations above 1% are not recommended. 500ul $104.42 from VWR

All in all for regular genotyping PCR which is most of what we do the GelRed works very well and while it's somewhat more expensive than the other dyes, it is a substantial difference in sensitivity. In the end the decision comes to whether you want to have an EtBr free gel bench and how much money you are going to spend for the more expensive dye.
5g of EtBr ($64.20) will make 1L of 10,000X solution (final concentration 5mg/mL), while 1L of GelRed at 30,000X would cost $35,000. Yet, 1L of stock will make 100,000 100mL gels: if you run 10 100mL gels a week or around 500 gels a year, you'll spend $0.32 in EtBr or $417.68 in GelRed. I would be interested in finding out how much you spend yearly for EtBr disposal.