recommended pcr kits? - (Nov/10/2005 )
hi everyone,
i've been having trouble getting a genotyping PCR to work, and I've been thinking about switching to a kit. What are some of the better PCR kits out there if one is simply interested in reliable genotyping results? Thanks.
logan
KOD from Novagen is the bomb
it's 'spensive, though; may not be best for your needs
i think my problem may be my tissue prep prior to the PCR. I've been using Proteinase K, but it's been very erratic, and I'm wondering if there are any good tissue prep kits on the market that might help me out? thanks.
logan
HI
if you have trouble in running your PCR, you may contact our labs at madurai in the following address for procurement of kits.
oncophyta labs
2/118 palkalai nagar East
Madurai-620021
Ph:091- 0452-2459586
094432 58228
Contact: Dr.G.Shanmugham
e.mail: shan1938@yahoo.com, oncophyta@yahoo.com
there are people in my lab who have tried PCR kits - and found that ultimately they are just not worth it - if u are going to spend the money i'd say get a better tissue prep kit, and do PCR the good old way.
if u are having problems getting a good quality template, then try out qiagen DNA extraction kits, they are very good, and affordable.........u have a kit specifically for extracting DNA from tissues, if u r specific about the species u will have them also....alternatively kits from life technologies called auprep is also good......the yield of DNA is good and even the quality of DNA......obtained.............u can also store them for considerably a long time.....
i hope this helps
good luck
i've been having trouble getting a genotyping PCR to work, and I've been thinking about switching to a kit. What are some of the better PCR kits out there if one is simply interested in reliable genotyping results? Thanks.
logan
For high-fidelity cloning and genotyping, I would select Phusion Polymerase (Finnzymes Oy, distributed by NEB: http://www.neb.com/nebecomm/products/productF-553.asp ) or Accuprime Pfx (Invitrogen). I use both of these with great success and no errors. Phusion is slightly cheaper and better because of enhanced processivity, IMHO.
For cheap screening, I use Promega GoTaq MM.
-Matt
Taq Biomix red is a good all in kit from Bioline but is low fidelity. Expang HIFI PCR from Roche is pretty good and is HI-fidelity. Also leaves a TA tail for TA taile cloning such as in TOPO. KOD is very good also but costs a bit
PCR kits aren't any better than the regular enzyme + buffer. I even prefer the regular enzyme so that I can (if needed) adjust Mg-concentration.
For reliable genotyping, go for expand hifi from roche, platinum taq or platinum/accuprime pfx from invitrogen, pfu turbo or ultra from stratagene, or phusion from fynnzymes (through NEB) or... or... (practically: ANY enzyme or enzyme mix with a higher fidelity than taq).