Magnetic beads: An alternative method to enzymatic purification for SNaPshot reactions

Abstract

This work reports the novel application of magnetic bead systems as an alternative to enzymatic purifications for SBE probes up to 100 bp. The Ampure and Cleanseq (Beckman Coulter Co.) systems allowed simpler and faster purification of both multiplex amplicons (evidenced by a very low noise to signal ratio) and fluorolabeled SBE probes (whose recovery results in high intensity peaks). A standardized and detailed protocol is presented for validation in small to medium forensics and population genetics laboratories workflow.

Steroidobacter gossypii sp. nov., isolated from cotton field soil

“…Briefly, the DNA sample was fragmented to a size of 350 bp by sonication and then DNA fragments were end-polished and ligated to paired-end adaptors for Illumina sequencing with further PCR amplification. PCR products were purified (AMPure XP system) [20], and the library quality was analysed using a bioanalyzer (Agilent 2100) [21]. The draft genome of S1-65 T was sequenced using Illumina HiSeq 4000 PE150 apparatus [20,22] at Tsinke Bioinformatics Technology (Beijing, PR China), and assembled using SOAPdenovo [23], SPAdes [24] and Abyss software [25], respectively.…”

“…PCR products were purified (AMPure XP system) [20], and the library quality was analysed using a bioanalyzer (Agilent 2100) [21]. The draft genome of S1-65 T was sequenced using Illumina HiSeq 4000 PE150 apparatus [20,22] at Tsinke Bioinformatics Technology (Beijing, PR China), and assembled using SOAPdenovo [23], SPAdes [24] and Abyss software [25], respectively. The assembled results from the three assemblers were further integrated with cisa software [26].…”