RNA extracted from liver tissue, here is the gel electrophoresis for that sample. first column is the RNA marker, would anyone please let me know what do you think of those rna sample?
Hi Li,
For future questions, it's good to add size markers to gel pictures and some info about the samples - it makes it easier to interpret results.
From that gel picture, I'd say your sample 1 and 2 were unsuccessful for RNA extraction but 3 and 4 were successful. The bands in samples 3 and 4 are the right pattern or fingerprint for ribosomal RNA - I've annotated your gel with the band names, using eukaryotes as an example - though the same pattern applies for prokaryotes.
That high molecular weight band present in all four samples may be gDNA, based on where the band is sitting in comparison to the rRNA bands. Do you use DNAse as part of your protocol?
Gel pictures can give you a good idea about integrity of RNA, though I'd highly recommend using Bioanalyzer to check quality and quantity of your extracts.
All the best,
Anna Liza
Hi Li,
For future questions, it's good to add size markers to gel pictures and some info about the samples - it makes it easier to interpret results.
From that gel picture, I'd say your sample 1 and 2 were unsuccessful for RNA extraction but 3 and 4 were successful. The bands in samples 3 and 4 are the right pattern or fingerprint for ribosomal RNA - I've annotated your gel with the band names, using eukaryotes as an example - though the same pattern applies for prokaryotes.
That high molecular weight band present in all four samples may be gDNA, based on where the band is sitting in comparison to the rRNA bands. Do you use DNAse as part of your protocol?
Gel pictures can give you a good idea about integrity of RNA, though I'd highly recommend using Bioanalyzer to check quality and quantity of your extracts.
All the best,
Anna Liza
Your RNA samples aren't so good, but depend for what do you to use. For cuantitative expression gene, I don't think this samples can you serve. The integrity in gel also depend about the concentration of sample. Regards from Mexico.
Hi , out of interest - my answer got down voted. If anyone is happy to get into that discussion, I'd love to know why - this is a pretty integral part of my PhD and RNA extractions are being a pain. If I wrote something that was incorrect I'd like to know so I can improve my own work!
Hi Anna Kretzschmar, When I saw the question yesterday - thought of answering, but you already answered it correctly. I was also surprised to see that somebody down voted it. Again I have gone through the answer and find nothing wrong in it, thus up-voted. I recently submitted my PhD thesis and during my PhD work I extracted RNA almost from 800 samples (mammalian Cell and Tissue both) for microarray and qRTPCR. So I am pretty experienced working with RNA and also would like to know what is wrong in your answer too.
I agree completely with Anna´s answer. The RNA looks just fine and, although it might not be suitable for quantitative procedures (personally, I think it would be fine), It´s good enough for library construction and/or fishing out some cDNA.
Cheers from Brazil
pper most band is of gDNA. Try to refine ur Extraction protocol. U will get only 3 bands of RNA if u will get good purity
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