The reproducibility of AFLP depends on many factors, and I would like to ask on the claims that may be required for optimizing the reaction, or how to optimize the AFLP reaction?
Hi Mohamed,
The "reproducibility" of AFLP, do you mean to obtain the same pattern of bands in different times?
Dear Canwei,
Yes, I mean with the reproducibility of AFLP to obtain the same pattern of bands in different times. Thanks for your interest!
Dear Mohamed,
What kind of chemicals do you use? Do you use ready to use kit or inhouse chemicals? The most important points are DNAs and components. I suggest you to use ready to use kits and high quality DNAs.
Dear Dr. Sigva,
Thanks a lot for your response! I'm using CTAB method for DNA extraction, and precipitating DNA with isopropanol, but I faced many problems with the quality or the quantity of DNA with a plant rich in polyphenols and polysaccharides, may be one can try Kits but I have heard that Kits are causing more fragmentation for DNA that lead to smears during running especially I depend on gel electrophoresis in estimation DNA concentration. For optimizing the reaction, which kits or method you recommend?
Dear Mohammed,
In house ctab method is the easiest and cost effective method for dna isolation, as you mentioned about, you can face many problem. I suggest that you can use dna isolation kits. When i work in AFLP, i used DNeasy plant mini kit from Qiagen. It is really good isolation kit. After isolation you need to measure your dna quantity and quality and you should arrange your dna (this is very crucial step). For AFLP process, i used AFLP® Starter Primer Kit from life technologies. My opinion, if you can get high quality dna and start desired amount of dna, you cannot face any problem during the all process.
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