Ours is new lab situated in Rural Medical School in Southern India and will be operational from May 2015 .I have very little experience in PCR. Kindly suggest with a list of essential chemicals for PCR from start to end . I plan to do more of molecular epidemiology work in human viruses and plan for about 1000 - 200 rxns per year. Thanks
for pcr necessary buffers mgcl2 enzymes and basic things for pcr reaction can be obtained NEB. Other than that you need agarose, etbr, tae or tbe buffer components for running agarose electrophoresis. If you going to work on expression then you might need vectors depending upon your work, restriction enzymes depend on your vector, ligase, sds, APS, acrylamide, bis acrylamide, PBS, IPTG, competent cells, and many..
Give clear details that what kind of work you going to do in your lab.
I agree with the previous replies and would kindly ask you to explain the PCR you are planning to perform.
Depending on the type of PCR (what is your target gene?) your are planning and the type of sample (human tissue, soil, marine, ...) I would also recommend to use BSA (Bovine Serum Albumin) or DMSO (Dimethyl sulfoxide). Of course you would also need the "regular" PCR chemicals which includes the polymerase, buffer, H2O, dNTPs and primer (as mentioned by the others).
Dear Anna Kopf, Elizabeth Pavez Lorie and Pankaj K. Bagul
Our lab will be doing mostly RT PCR and samples could be of human source like plasma, serum and stool. we will working with viral RNA or DNA. We will use kit based extraction methods for nucleic acids and also master mixes. I would like to have baisc chemical list for PCR.
Basic PCR components and chemicals:
1. DNAse-free water
2. Primers
3. dNTPs
4. MgCl2
5. DNA polymerase (different choice: regular, long-ranged, hot-start, High-Fidelity.... depending on your need)
6. DNA template
7. PCR buffer
8. PCR additive, such as DMSO, BSA etc. (in case your DNA template is GC rich)
How many PCR types and what are they do? can you enlighten in this way?
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