Can someone motivate the use of 1M beta mercaptoethanol (7.5% v/v) in HMW plant genomic DNA prep?
I understand the need for BME to reduce polyphenols and keep reduction state but the '1M' used in my protocol seems absurdly high to me and generates a large amount of toxic waste (and may be uselessly toxic for the user).
The buffer also contains PVP-10, spermine and spermidine as the original and related IB buffer published by Simkova et al (2003) but much more BME than the 15-45mM reported in the paper.
Thanks in advance
I feel that mercapto-ethanol has a role in protein denaturation as well besides the removal of polyphenols, to break disulfide linkages. I am not sure, but maybe a higher concentration is required to break the disulfide linkages in the protein.
seems very high to me as well. Not sure what is the final use of your DNA; in our lab we routinely use a CTAB extraction buffer containing 0.2% B-mercaptoethanol and gives a very clean DNA in terms of A260/230 and A260/280. We work with Setaria, rice, arabidopsis, tobacco etc. and use it mostly for PCR.
2-Mercaptoethanol use for denaturation of proteins (HMW) reducing disulfide bonds in protein molecules. Also use in procedures of RNA isolation remove enzyme ribonuclease during cell lysis.
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