If its in a laboratory, we can use any modern technique to get DNA from plants without any damage, but what about labs in remote areas which don't have well equipped laboratories? Are there any simple methods to isolate DNA for genetic analysis?
Hi,
I am using a DNA extraction kit from Epicentre (Master Pure DNA purification kit).
The only thing you need is a centrifuge and a water bath (or a Thermomixer).
Cheers, Nadine
HI Abhi,
Back in grad school we used the Dellaporta technique for extracting DNA from many plants and it gives high quality DNA that amplifies by PCR without any trouble. However, for the few plants whose leaves have excessive amounts of sticky mucilagenous material, the Dellaporta DNA extract was troublesome in PCR. Reprecipitation helped sometimes but not always. What definitely worked, in all cases, was the Qiagen DNAEasy extraction kit, but we did not have the funding to purchase many of these kits to process the large number of pant samples we studied. So we used the Dellaporta routinely and for the few troublesome mucilagenous plants we used the DNAEasy kit. The dellaporta protocol may be found at
http://www.arabidopsis.org/comguide/chap_3_dna_techniques/5_modified_DNA_miniprep.html
I recall that the lab next to ours used the CTAB method to extract DNA from cotton and citrus leaves. The CTAB protocol is here
http://www.cilr.uq.edu.au/UserImages/File/Plant%20Genomic%20DNA%20Extraction%20by%20CTAB%20_2__Fiona.pdf
Best Wishes
Here's another one that pretty much sounds like what you are looking for. I think we used something similar for public demonstrations. There are also protocols available to isolate DNA from plant tissue using laundry detergent or dish liquid but you will always need a centrifuge (or a drilling machine with some self-made support and very stable safety glasses)
Also possible to try DNA extraction with CTAB (Cetyltrimethylammonium bromide) (See Doyle and Doyle, 1987). It is old protocol, but it successfully work in most cases and needn't some special equipment or reagents and easy to use.
And in general the choose of DNA isolation methods greatly depends on for what analysis this DNA will be used in future.
Best wishes
Nowdays, there are direct PCR kits which did not require DNA isolation. Could be a good option.
The Qiagen Plant DNeasy Kit is easy to use. http://www.qiagen.com/products/genomicdnastabilizationpurification/dneasyplantsystem/dneasyplantminikit.aspx#Tabs=t0
Doyle and Doyle method is good I belief but sometimes u need to bring some modification according to species. Here u need only a centrifuge and water bath.
The Isolation of pure, intact and high quality DNA is very crucial for any molecular studies. However, The chemotypic heterogeneity among various plants may not permit optimal DNA yields with a single protocol. Therefore the extraction protocol may vary from plants to plants. So here question comes, which plant you are really interested in. I strongly agree with the previous suggestions by the experts. I too have standardized DNA extraction protocol for plants for plants harbouring high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. I have attached the file for your kind perusal or u can follow (https://www.researchgate.net/publication/233419566_DNA_Extraction_Protocol_for_Plants_with_High_Levels_of_Secondary_Metabolites_and_Polysaccharides_without_Using_Liquid_Nitrogen_and_Phenol?ev=prf_pub)
Article DNA Extraction Protocol for Plants with High Levels of Secon...
thanks a lot for your info,
i m voluntarily teaching in govt schools which is in buffer zone of western ghats, so students in 10+2 are very much interested in extracting plant DNA,
this will help a lot, thank you again ,, :-)
Sunil Kumar Sahu I would like to use your protocol but I have one question. I do not understand step (V). Do I need transfer upper phase (separate the plant material) after centrifugation and then add extration buffer? It is not clear for me...
Thank you in advance!
Dear Dunja Samec, Sorry for the late response. I am glad to know you are interested in using my protocol. let me make you more clear, in this protocol I have used two different buffers namely suspension buffer and extraction buffer. After adding suspension buffer followed by incubation, u have to discard the supernatant and extraction buffer is to be added to the pellet. In the next step after centrifuge you have to take the upper aqueous phase and transfer into new vial. Then you can proceed for further extraction process. Hope I am clear to you. If still you have doubts please feel free to contact.
I had a dilemma if I need discard supernatant after suspension buffer or not. Thank you for answer!
We have published and excellent protocol, fast, cheap, efficient and created to obtain high quality and purity DNA. Barra et al. 2012. Simple and robust DNA extraction method for the large-scale analysis
of genotypes containing high polyphenolic content, such as landraces
of Solanum tuberosum and Zea mays, Cien. Inv. Agr. 39(3):593-601. 2012
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