Is there any DNA extraction kit (affordable and reliable) available in India to extract DNA from minute bryophyte like Cololejeunea ( using 2-3 plants only) which in grams will be less than 2 mg. It will be really helpful to me if it is possible. Any suggestion for the methodology is welcome.
I suggest you to contact Qiagen, because if they have not specific kits for the extraction of DNA from minute plant material, they have protocols or kits for contcentrating the result from more than one extraction. Anyway, they have good staff for the resolution of your problem.
Best regards and good luck!
Maybe you can try QIAamp® DNA Micro Handbook (qiagen) that was developed for tissue sample of less than 10 mg.
It works on very small pieces of poplar roots
According to our experience even small epiphyllous liverworts can be used for DNA extraction, but extreme care should be practiced to avoid contaminating with fragments of other species living on the same leaf.
In bryophytes phylogeny study, most of the researcher use the kit of Qiagen to extract DNA, especially in epiphyllous liverworts, for they are so tiny. And in my experience (sample of moss), 2 mg pure dry sample is enough. Usually I use CTAB (Doyle & Doyle,1987) to extract DNA , which is a method that most protocols can be changed according to your sample. The most important thing is the quality not the quantity of the sample, the year of the sample (better in 10 years from now) and the condition of the sample (e.g. without fungi or algae, keep dry in desiccant, keep in a low temperature). The last but the most important thing is avoiding other species like Dr. Pócs said.
Best wishes!
Thank U all for your valuable suggestions. I feel it is not so simple to me as Mr Manoj suggested. For minute plants, separate kit is needed. Today we have attempted using a kit with about 20 mg of other bryophyte but not the minute plant and DNA.
The protocol provided with kit suggest to use 10 mg and above sample which is posing difficulty in dealing with plant having less quantity specially the members of lejeuneaceae. Therefore, i am looking for a suitable kit and protocol for that.
with regards.
Dear Dr. Singh, after extract DNA, you should test the quality and quantity of them using a normal DNA spectrophotometer, and try a PCR to test the quality of DNA, not all the DNA extracted can do PCR well. Some of them maybe breakdown into very short fragments。
Dear Dr Yu
Still I am facing similar problem. I have attempted the extraction with CTAB (Doyle & Doyle,1987) and SDS and Nuceospin mini plant kit but no DNA could be extracted for less quantity material (I used even 15 mg), no gel image appears. Only i could extracted where material are more than 100 mg. I will be very thankful if you can suggest any changes in protocol of extraction so that i can deal with materials of lesser quantity.
thanks for any help.
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