I used to synthesize cDNA from the RNA, extracted from the leaves by TRIZOL method. Im not getting a good 260/280 as well as 260/230 values. Should i proceed for further experiments?
I experienced using TRIZol and other DNA extraction kits from QIAGEN. TRIZOL can give you a good yield of RNA but with a lot of contaminants of both DNA and protein, despite you can use a DNASE or Proteinase treatments to increase the quality.
You can check the RNA quality using formaldehyde agarose gel electrophoresis and based on that you can decide wether you need to use DNase treatment or not.
Karima Ben Mansour and Mohammed Elwetidy thank you.. but my question is, should i proceed further?
As i said the quality of RNA is only visualized by gel electrophoresis, if you did not see any smearing and the 2:1 ratio was fine then you can proceed to cDNA. However, what i can advice you to do in order to improve the ratio is to re-precipitate RNA and reapeat washing step using 75 % Ethanol then measure it again. . But, if you will use your cDNA later for qPCR then i recommend you to do a DNase treatment.
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