I have a problem with isolation of genomic DNA from medicinal plants high in secondary metabolites. Actually, I get some DNA (i could measure on nanodrop) and I see DNA on agarose gel but my PCR reaction failed. PCR reaction work with other samples and all primers. I think that something from DNA tamplet bothered reaction. I have air-dried samples of problematic medicinal plant. Does anybody have some recommendation?
I tried CTAB method (all variations) and commercially available kits.
Hello,
i agree with jyoti, but slightly varies and simple procedure this too u may try. 1.Buffer(pH 9)- 50mM Tris-cl, 100mM EDTA, 200mM NaCl
2.0.3M sodium acetate
it was ground with 0.5ml buffer. The homogenate ia added then, 30µl of 20% SDS is added. The tubes are inverted for few times. Phenol: Chloroform in the ratio (1:1) 400µl:400µl is added. the tubes are inverted several times. they are centrifuged at 15000rpm at 4oC for 15 min. The aqueous phase is collected to that 100µl 0f 0.3 M sodium acetate is added. Then 0.8 volume ethanol is added. The tubes inverted at placed at -20oC overnight. The tubes are centrifuged at 15,000 rpm for 15 min at 4oC to collect the pellet. The pellet is washed with 70% ethanol. After drying the pellet is washed, dissolved in TE buffer (pH8) and stored at -80oC.
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