I would possibly like help in extracting RNA from pigeon pea leaves. I am currently using the Maxwell 16 Lev plant RNA extraction kit. I have conducted 2 extractions yet both yielding low concentration (+/- 50 ng/ul) and purity (1.4- 1.6). I also treated my cDNA with thermofishers DNase treatment kit yet although the purity improves, I am still unable to visualise my samples on a 1% gel. I plan on synthesizing cDNA from this RNA and cloning into pGEMT Easy afterwards.
I am all ears for any possible troubleshooting.
We have been using phenol extraction method for student practicals
Try adding 3% of β-Mercaptoethanol in the lysis buffer to reduce rnases activity.
check the date on your kit, they do expire after about 1 year. Is this a column kit a pellet kit? If your sample has lots of secondary compounds (resins etc.) you may need to decrease the amount of starting material.
Simply try with the TRizol (invitrogen) reagent. Works best for plants., There was also a protocol form S. Korban I guess if I remember correctly from my back days...
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