Taq polymerase expressing E. coli requested - (Jul/25/2008 )
Hi everyone I'm new here,
I'm a poor medicine student (5th year) with alot of spare time this summer working on a rich laboratory for a rich uninterested professor who doesn't want to support my out of the square thinking experiments (perhaps he is right). We are routinely doing PCR with kits from Roche et al for the patients at our hospital. Now, my professor is not interested in saving money for the hospital and ultimately for the taxpayer, but i am. So I took a tour around our lab to look for equipment needed in purifying proteins and I found plenty: chromatography columns, agarose, an old UV-spectrophotometer etc etc.
My plan? To mass produce Taq polymerase for the lab. Now my professor does not want to know anything of my idea but i am allowed to use the laboratory glassware and chemicals. I have gathered the equipment needed but lack the money and connections to obtain a strain of E. coli that expresses Taq. Is there a friendly soul out there that could help me out by sending me a small culture of this wonderful bacterium?
Best Regards
Chocolate agar
"1 year to go"
Hi Chocolateagar,
maybe this is not the best way to spend your summer-time 
and to save money...
If you think about the costs of Taq or a hot-start Taq in an experiment and all the other components which are needed for an experiment (plastics, time, dNTPS, buffer, preparation, pre- and post-pcr experiments) I think it is not worth to spend your time in doing this. Especially if you do not want your collegues to shout at you 
if one of their experiments would fail with "your" Taq. It is not that easy to purify the Taq and make it DNA-free (and DNase free) which is very crucial for successful specific PCR. How do want to quality control your Taq and how to determine the activity compared to commercial ones?
Maybe it would be better to relax during your free time, help your collegues with their work or establish a new helpful or time-saving technique in your lab... Or you think about where your lab can save "real" money beside Taq - maybe use another kit for DNA purification (Qiagen is very expensive) and validate the new kit in the lab workflow. I think, if you carefully think again there are many better projects you could spend your time with 
and help your group, the patients and tax-payers and your knowledge
Let me know how you finally decided
maybe this is not the best way to spend your summer-time
and to save money... If you think about the costs of Taq or a hot-start Taq in an experiment and all the other components which are needed for an experiment (plastics, time, dNTPS, buffer, preparation, pre- and post-pcr experiments) I think it is not worth to spend your time in doing this. Especially if you do not want your collegues to shout at you
if one of their experiments would fail with "your" Taq. It is not that easy to purify the Taq and make it DNA-free (and DNase free) which is very crucial for successful specific PCR. How do want to quality control your Taq and how to determine the activity compared to commercial ones?Maybe it would be better to relax during your free time, help your collegues with their work or establish a new helpful or time-saving technique in your lab... Or you think about where your lab can save "real" money beside Taq - maybe use another kit for DNA purification (Qiagen is very expensive) and validate the new kit in the lab workflow. I think, if you carefully think again there are many better projects you could spend your time with
and help your group, the patients and tax-payers and your knowledge Let me know how you finally decided
Oh my, are you my Professor?
Jokes aside, you have a point The Professor. It will be timeconsuming for me, very timeconsuming. The thing is, I'm born to work at the laboratory. Weighing, measuring, pipetting, western-/northern blotting, pcr, chromatography, HPLC, documenting, searching, pubmed... you name it. I wouldn't mind having this as an alternative project running beside the main one at our lab (if it works, all the better, if not, i have to look at my protocols). If one wants, there is time for everything. It's just a question of priority.
To be frank, what's bothering me the most, apart from trying to be the hero of the taxpayer, is that all the skill and if you want "art" is disappearing. It's just kits today. DNA extraction kit, purification kit, electrophoresis kit, Roche kits, Qiagen kits, Becton & Dickinson kits, Molzym kits...yes they are making life easier and safer at work but I would like to know the details, and I would like to have worked through it all before I go through the kits. I know you are a friend of kits, The Professor, and i see the benefits of these. (And another thing, in my opinion, the companies are way overcharging us for their work, but somehow, for a scientist, it is often still "more profitable" to buy labor time -in the form of a kit - from a companies, than to hire a full time technical assistant to do the job.)
I am 27 and I'm being asked not to ask any questions, or if so, go read it at the local library. "Hey man, relax! You don't have to know everything! You will go crazy if you know everything! Why do you want to waste your time on what has already been done?". While others prefer fishing, hunting, sports, movies, games, music, politics, philosophy, i prefer lab work. To me THIS is leasure time, I simply enjoy this... apart from spending time with my fiance of course. It's hard to tell others this without being laughed at, so internet is the only way to go.
Please forgive my rantings and my misspellings
Chocolateagar
"1 more year..."