I extracted genomic DNA and total RNA from fruiting bodies and mycelium of basidiomycota in a daily routine. Some pigments can not be remove from DNA/RNA with column. Consequently, DNA/RNA quality show low A260/A230 ratio. Does anybody know a good removal method of the pigments in the genomic DNA from the fruiting bodies (mushroom)?
What method did you use to DNA and RNA extraction? Did you use kit or manual extraction? In my case DNeasy and RNeasy plant kits works well. If I have problem with DNA quality I change methodology to manual and use protocols with CTAB.
Thank you for your response, Dr.Adam. I use RNeasy Plant mini kits (Qiagen) for RNA extraction. This kit works well in my lab, too. However, I am not satisfied DNeasy because it is low yeild. So, I extracted gDNA from mycelium using by modified ISOPLANT II kit (Nippon gene, Japan) . ISOPLANT II Kit including benzyl chloride. But ISOPLANT II Kit is not effective pigment removal from fruitng bodies. I tried CTAB method, but failed. Provably, ethanol prep. in each method is bad step. Some pigments are precipitated with gDNA by ethanol prep. The pigment could not be removed in downstream of ethanol prep. step.
Maybe this article will help:
Islas-Flores, I., Peraza-Echeverría, L., Canto-Canché, B., & Rodríguez-García, C. M. (2006). Extraction of high-quality, melanin-free RNA from Mycosphaerella fijiensis for cDNA preparation. Molecular biotechnology, 34(1), 45-50.
I wish to express my strong appreciation to you for introducing paper. It'll be useful.
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