Ryan Ho

Hey everyone,

In my spare time, while I'm taking a break from my experiments not working, I'm a volunteer for the Association of African Entrepreneurs (AAE), a charity that helps promote and support entrepreneurship throughout Africa. Our current goal is to raise $4000 from 50 unique donors by September 30/2012 at which point we would obtain permanent status on the GlobalGiving website that would set us up for long term funding. We only have until that deadline to do it, so any help would be great! GlobalGiving is an organization that connects donors to projects in developing countries, so this is a great opportunity for us to expand and connect to more entrepreneurs. Click here to make a secure donation: http://www.globalgiv...ts/project-aae/

This is a really great cause since we are investing in African entrepreneurship, rather than getting people to rely on the charity. I assure you this is 100% legitimate, you can verify with the sites given below.



If you want to look into it more here are some helpful websites.

http://www.globalgiv...ts/project-aae/
-    AAE project web page on the global giving site. Access this page to make a donation to AAE

http://www.globalgiving.org/
-    The global giving website, which is hosting the web page for donations to AAE

http://en.wikipedia....ki/GlobalGiving
-    Information on the global giving website

http://www.aaeafrica.org/projectaae/
-    Information about the Association of African Entrepreneurs the charity that is being supported by donations



Thanks a lot for your help!

I'll allow it, and leave it here for now, but may move it to another subforum later - bob.

Alright I'm going into my 2nd year of grad school and for one of my experiments I'm doing a microarray on swarming cells in P. aeruginosa. Swarming is a type of motility in P. aeruginosa, and I'm doing a microarray of the wild type vs. a mutant which hyper swarms. I've had to work out some problems but I finally got the phenotypes I wanted. Basically I do everything up to and including the genomic DNA check after RNA extraction and DNase treatment, at which point a technician takes over and does the microarray.

I've attempted this twice, but each time there was too much variability between each of my biological repeats (P-values were too high) and this has eaten up ~8 months of my time. Are these types of problems normal? I feel like such a straggler right now but I'm still trying. I'm on my 3rd attempt, and I really have no idea what's going on. Here's a basic outline of what I do,
- Inoculate my swarm plates and let my mutants swarm for 18 hours
- Collect the swarm cells with a swab and "add" the cells to water + buffer by swishing the swab in the solution
- Add 2 volumes of RNA protect
- Centrifuge for appropriate amount of time
- RNA extract using QIAGEN RNA extraction kit
- DNase treat with Ambion DNase1
- Genomic check w/ rpsL

I've made sure all the conditions were the same, each BR was treated exactly the same to the best of my ability. I really don't know what's going on, so any help/suggestions would be helpful.

Thank you