I have to isolate more than 2500 wheat leaf samples for screening. Even we use robotic system, dna isolation is very time consuming and costly. We will use only 2 primer pair for screening. I wish to perform directly PCR by skipping DNA isolation. Phire Plant Direct PCR kit is also available but not cheap. If any alternative to the kit, I will try it.
Thanks in advance.
Hi Fatma
I have tried a manual method for direct PCR on different plant species and it worked well. You just need to take a little piece (2-3 cm) of plant young leave, homogenise it in sterile 20 to 50 ul water. I did it with the help of tissue homogeniser with microcentrifuge tube pestle. Use 2-3 ul of this extract as template and run the PCR with standard conditions (I took the caution to keep the extract on ice till further use). I have also tried MOLECULE-ON, New Zealand Direct Plant PCR Kit and also work well.
All the best.
Dear Fatma Akın
Pls. find the attached files
https://www.ncbi.nlm.nih.gov/pubmed/23051689
https://www.kapabiosystems.com/product-applications/products/pcr-2/kapa3g-plant-pcr-kits/
http://www.seoulin.co.kr/dm/catalogue/thermo/thermo_mb/directpcr/direct_pcr.pdf
https://www.jove.com/video/3844/genotyping-of-plant-and-animal-samples-without-prior-dna-purification
Hoping this will be helpful
Regards
Prof. Houda Kawas
Hi Fatma
Did you ever try the method "
A rapid and inexpensive method for the direct PCR amplification of DNA from plants
Article in American Journal of Botany 97(7):e65-8 · July 2010"
I tried it in my lab recently. And it is good for some plant species. Hope it also works for your materials.
Zha
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