![]() Usually DNA, mRNA, and small RNA sequencing libraries are constructed with distinct linkers and/or chemistry, forcing customers Although this strategy significantly decreases the sequencing cost, it cannot solve the problem that the overall libraryĬonstruction cost using commercial kits could easily surpass the sequencing cost. Individual samples are usually cloned with specific barcodes, pooled, and sequenced as a single library for cost-sharing,Īnd then debarcoded to obtain sample-specific sequences ( Stiller et al. Since a single sequencing run is usually sufficient for analyzing multiple samples, In this technique will have a broad impact. Over the past decade, it is becoming a standard tool for gene analyses in biomedical fields. High-throughput sequencing has become a revolutionary technique for analyzing DNA/RNA. Previous Section Next Section INTRODUCTION constructing sequencing libraries without gel purification.high-throughput or next-generation sequencing.Moreover, the all-liquid-based reaction can be performed in ![]() Not only is our method more convenient for cloning modified RNA than available methods,īut it is also more sensitive, versatile, and cost-effective. Moreover, this method can also clone mRNA, simplifying the need to prepare twoĬloning systems for small RNA and mRNA the barcoded PCR primers are also compatible with non-cDNA cloning applications, including Theħ-h cloning process only needs ∼1 h of labor. Unmodified small RNA in an all-liquid-based reaction carried out in a single PCR tube with as little as 20 ng total RNA. Here we present a new strategy to clone 5′ modified or It is difficult to clone small RNA with a small amount of total RNA. Unlike mRNA, small RNA often contains modifications including 5′ cap or triphosphateĪnd 2′- O-methyl, requiring additional processing steps before linker additions during cloning processes due to low expression levels, ![]() This method usually needs a cDNA/DNA library High-throughput sequencing has become a standard tool for analyzing RNA and DNA. ![]()
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