I am Nicholas from the University of Malaya. I have been performing RNA extraction from whole zebrafish larvae for some time using NucleoZOL as my lysis buffer and passing my samples through column tubes according to manufacturer's protocol. Typically, I achieve yields of >100 ng/μl with high purity from a pool of 30 larvae.
My current project requires RNA extraction from the brain and gut of 7-day-post-fertilization (dpf) zebrafish larvae. Despite various sample sizes (30, 60, and 150 larvae per sample), I have encountered difficulties in RNA extraction using the same method. Recently, I purchased a microcentrifuge pestle to assist in homogenization, but I am facing challenges as debris tends to stick to the walls of the microcentrifuge tube. I'm concerned about potential RNA degradation during the process, especially since I did not add beta-mercaptolethanol during extraction.
The highest yield I obtained was 10.7 ng/μl (n=30), while yields typically range between 0.5-3.7 ng/μl regardless of sample size. I am seeking advice on how to improve RNA yield and would greatly appreciate any tips or suggestions, particularly interested in any methods or adjustments that could enhance RNA extraction efficiency. Thank you.
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