![]() ![]() The DNA library construction strategy is straightforward, basically ligating fragmented-and-end-fixed target DNA with partiallyĭouble-stranded DNA linkers, as used by commercial kits ( Illumina 2012 Kircher et al. Which are then used to design primers for amplifying and reading sequences/barcodes ( Goodwin et al. In most sequencing platforms, a DNA or cDNA library is made by ligating fragmented DNA/RNA with platform-specific linkers, If a laboratory wants to prepare its own barcoded PCR primers, Primers for obtaining the final amplicons are also different. Since the linkers for DNA, mRNA, and RNA libraries are different, the corresponding PCR Most commercial kits only provide just enough primers for a specified number of library constructions, allowing no Lack of compatibility among these expensive kits and transparency for the protocol details often lead to confusion and To purchase different kits ( Reuter et al. 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. ![]()
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