Monday July 17, 2006: Chulalongkorn University

Today was our first day in the lab at Chulalongkorn University. Sunee met us in the lobby of our hotel at 9:15am and walked us to the hospital, about 10 minutes away. A conference room was waiting for us, along with the other employees of this Thai lab. Kiat, the director gave a quick presentation of the data they had gathered thus far. Next, Todd presented an overview of what was expected from each site for the contract (Peru, Thailand, China, as well as Boston's role) by the end of this month, the end of year three for Contract II. There's a LOT to do in Thailand! Todd included a list of more specific duties that need to be worked out while Liz and I are here. This should really help Thailand get up to speed with their part of the Contract.
My group, Sven (PhD) and Mew (technician) gathered after the meeting in the conference room and brought me around the microbiology and cellular labs. The facility is 2 years old, and they have ample space. Everything is very white and appears clean. We gathered for lunch in the conference room and had Thai take-out which was very delicious. The fresh fruit was quite abundant as well.
After lunch I put my team to work. We isolated viral RNA from two HIV+ plasma samples using a 2 hour spin and also a regular extraction- both with the Qiagen vRNA mini-kit. We then used 3 dilutions (1:1, 1:10, 1:100) of each isolation and did 12 RT-PCR reactions with the Titan One-Tube enzyme. Mew, who did all the work, seemed like I had asked a lot from her, and 12 reactions being uite a small number from my point of view compared to the caliber of reactions that we do in Boston, was quite surprising to me. I found out shortly after the extraction, that they have individual reaction tubes so it takes a very long time to label each tube, hence the problem with doing many reactions at once. It also takes about 2 weeks to a month to receive reagents in their lab, while it takes as little as one day to four days to receive reagents in Boston. Also, they don't plan too far ahead, so when they are out of reagents, they order more and just wait until it arrives. Extremely inefficient, but they also can't afford to waste reagents if they don't get used. The labelling of each individual PCR tube slows things down tremendously, and insn't conducive to setting up many reactions at once. I'll see what I can do to help out the productivity...
The TC (tissue culture) room we use has no air-flow, no A/C. With a gown on, 2 hoods ventilating their exhaust, as well as a fridge and centrifuge, the temperature was well over 100 degrees F!
Liz's TC room was quite nice; actually, it was too cold! They leave their sandals outside and put on slippers to enter the TC room. There is a large office to the side of the TC room that has huge windows, a viewing room into the TC area. Her group, Man and Am also seemed to be asked to do quite a lot, especially since we each made them stay til 5:30pm.
Hopefully without the high spin and with the dilutions we used, we will find a good amount of template that will work. Right now, a 2 hour spin and 15ul of vRNA is way too much template I think, causing a large smear on their gels. If this works, I will have earned some trust and respect and should I make more adjustments to their protocol in the future, I think they will be sure to take my advice! Also, this could solve their problem immediately, and that'd be marvelous for all involved, not to mention saving them 2 hours right off the bat!