Plate Scale Tn5 based tagmentation library prep protocol v1
crossref(2024)
摘要
This protocol is a fork of (dx.doi.org/10.17504/protocols.io.bv5gn83w), which a demonstrated efficient and high-throughput tagmentation sequencing library prep protocol based on Picelli et al 2014. In this version, the tagmentation and amplification steps remain largely the same - based on the Tn5 transposase and KAPA HiFi kit, respectively. Here, significant modifications have been made to the library pooling and cleanup processes. Specifically, this protocol involves pooling the crude PCR products, followed by the removal of small fragments via a spin column kit, and depletion of large fragments through agarose gel extraction. These modifications enable faster processing times while still producing high quality sequencing libraries. This protocol has been optimized for sequencing Drosophila cell culture samples, from which high-quality genomic DNA (gDNA) can be readily obtained. The accessibility to high-quality gDNA simplifies the normalization of gDNA input amounts prior to tagmentation, thereby streamlining the pooling and cleanup steps.
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