What causes low 260/230 ratio for RNA & how to correct it - (Apr/29/2005 )
if your RNA is too concentrated, the ratio and purity will not accurate.
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
-sanjiun81-
QUOTE (sanjiun81 @ May 10 2007, 07:23 PM)
if your RNA is too concentrated, the ratio and purity will not accurate.
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
I agree with Sanjiun, if your RNA is too concentrated or too dilute then ratios can look very odd so try a few different dilutions
-auldmok-
QUOTE (sanjiun81 @ May 11 2007, 10:23 AM)
if your RNA is too concentrated, the ratio and purity will not accurate.
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
How about DNA?. is it the same issue as described above?, too concentrated lead to low DNA value?.
-quangtranho-
QUOTE (quangtranho @ May 14 2007, 10:25 AM)
QUOTE (sanjiun81 @ May 11 2007, 10:23 AM)
if your RNA is too concentrated, the ratio and purity will not accurate.
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
i had the same problems few weeks ago. my bioanalyzer's and nanodrop results like crazy. i try to extract so many times and thought my technique not right. but after somebody suggest to dilute my RNA, the results for nanodrop and bioanalyzer improved amazingly!!!
How about DNA?. is it the same issue as described above?, too concentrated lead to low DNA value?.
Yes -- you need to make sure your dilution is within the limits of detection of the equipment in order for your reading to be accurate. We make dilutions based on our expected yield, then redilute and reread if necessary. I've never used a nanodrop, but I know that on a spectramax, your OD260 should be somewhere between 0.2 and 2.0 to be reliable.
-MolBioGirl-
the pH of the buffer or dH2o that you used to elute your RNA may also affect the OD ratio I think.
-Stressguy-