Hello every one i want to know weather these bands i get for dsRNA is OK or not? my dsRNA concentration is about 2500-9500 each gene but bands. i have doubt need some kind suggestion also want to know how much UL i inject to insect cuticle and which part? picture attached with message thanks need suggestions?
You didn't mentioned the molecular weights of ladder you used. I hope it is a 2% agarose gel. Normally the isolated total RNA contains the higher percentage of ribosomal RNA (rRNA) what you see on gels. You cannot see the mRNA on gels due to its less concentration. Your gels contains three different samples, the third one is seems to be better with good concentration of RNA. You can proceed with this, but I don't have idea about insect biology. Your samples have higher amount of sheared genomic DNA, which is not good for further processing. Try to isolate good RNA with 2-3 bands visible on 2% agarose gel with no smear. Try to load less RNA (about 500-1000ng/well). Good luck for your future experiments.
Are you purified the dsRNA after DNase treatment, which used as template for invivo dsRNA synthesis.
Yes, DNAase treatment followed by running PCR with RNA as template could help.
Take help the protocol RNA isolation (RNeasy Mini Kit from https://www.qiagen.com) , which is quite useful.
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