Unlike mRNA, small RNA often contains modifications including 5′ cap or triphosphate and 2′- O-methyl, requiring additional processing steps before linker additions during cloning processes due to low expression levels, it is difficult to clone small RNA with a small amount of total RNA. This method usually needs a cDNA/DNA library ligated with specific 5′ and 3′ linkers. High-throughput sequencing has become a standard tool for analyzing RNA and DNA.
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