I extract DNA from different parts of dry herbs like dry leaves, seeds, stem, roots, seed coat, rhizome flowers, fruits etc via CTAB method and then I use that DNA for PCR amplification. I face many problems but the basic problem is that in many casrs DNA is not extracted at all and in some cases the extracted DNA is not pure in quality and hence does not give results in PCR. can anybody suggest any better method like extraction with spin column etc?
Hi dear Munawar! Working with that kind of plant material is always a high risk of not getting any DNA yield or PCR results. That's because DNA is so degraded that is not possible for PCR to induce amplification or is too much contaminated with polyphenols and other stress-dying molecules. Or it is in too low concetrations anyhow. Maybe try exploring highly sensitive DNA extraction and PCR amplification methods used in plant forensics methodologies? You can try google them on google scholar or any other academia platforms for papers and inputs about it. If you cannot find them also, try to google international/national institutions which are dealing with plant genetics or plant genetics and forensics (Germany, USA, China or similar countries).
Qiagen kits for plant and fungal DNA extraction might be also a good option but consult firstly with them or mentioned institutions, because, they are not cheep. I think there are some really sensitive DNA extraction methods which might help you with this. Hope that i was of some help.
The best a way for DNA extraction of Plant from fresh organs , the way with dry organs not efficient because the DNA exposed to damage and will be not clear
Dear Munawar Ali ,
are you using liquid N2 to break the cells? If not that could be another reason why you are not getting a result. We tried different DNA extraction kits and protocols to get DNA from Aquilaria spp. and were only successfull when using liquid N2. Hope this helps?
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