Rapid library preparation method

Manufactured by Roche

The Rapid Library Preparation Method is a laboratory equipment product offered by Roche. It provides a streamlined process for preparing samples for genomic analysis. The core function of this product is to enable efficient and rapid library preparation, a critical step in various genomic research applications.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (1 mentions) Superscript first strand synthesis system for rt pcr, by Thermo Fisher Scientific (1 mentions) Nebnext mrna second strand synthesis module, by New England Biolabs (1 mentions) Gs flx titanium sequencer, by Roche (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Experion automated electrophoresis system, by Bio-Rad (1 mentions)

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Rapid Library Preparation Method Manual (2 citations)

2 protocols using rapid library preparation method

Transcriptomics of Tarantula Venom Glands

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The tarantula spider, Ornithoctonus huwena, is not a protected species and found widely in the Guangxi Province of China. Three tarantula spiders were collected for study. No specific permission was required for these locations/activities. Venom glands of Ornithoctonus huwena were obtained two days after being milked via electrical stimulation, and ground to fine powder in liquid nitrogen. Total RNA was extracted with TRIzol (Invitrogen, Carlsbad, CA, USA) and used to construct a cDNA library. Full-length enriched double-stranded cDNA was synthesized from pooled total RNA using the SuperScript First-Strand Synthesis System for RT-PCR (Invitrogen) and NEBNext mRNA Second Strand Synthesis Module (NEB), according to the manufacturer’s protocol, and subsequently purified using the QIAquick PCR Purification Kit (Qiagen USA, Valencia, CA). The DNA library was prepared from 300 ng samples using the manufacturer’s instructions (Rapid Library Preparation Method, Roche). Sequencing was performed on a Roche GS FLX Titanium sequencer.

Zhang Y., Huang Y., He Q., Liu J., Luo J., Zhu L., Lu S., Huang P., Chen X., Zeng X, & Liang S. (2014). Toxin Diversity Revealed by a Transcriptomic Study of Ornithoctonus huwena. PLoS ONE, 9(6), e100682.

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RNA Isolation and Normalization for 454 Sequencing

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Total RNA from liver was isolated using Trizol Reagent (Ambion) according to the manufacturer's protocol. The integrity of the RNA was verified using an Experion Automated Electrophoresis system (Bio-Rad). cDNA synthesis and normalization were performed by the Evrogen Lab (Moscow, Russia). Total RNA samples were used for ds cDNA synthesis using a SMART approach (Zhu et al. 2001 (link)). SMART-prepared cDNAs were then normalized using the DSN normalization method (Zhulidov et al. 2004 (link)). The normalization procedure included cDNA denaturation/reassociation, treatment by duplex-specific nuclease (DSN) (Shagin et al. 2002 (link)) and amplification of the normalized fraction by PCR. One μg of each normalized cDNA sample was used to generate a tagged 454 sequencing library using MID adaptors following the Rapid Library Preparation Method from Roche. The libraries were sequenced on a GS FLX instrument using Titanium chemistry. Most libraries were sequenced in pairs in which two different libraries were pooled and loaded onto one sequencing region.

Mořkovský L., Janoušek V., Reif J., Rídl J., Pačes J., Choleva L., Janko K., Nachman M.W, & Reifová R. (2018). Genomic islands of differentiation in two songbird species reveal candidate genes for hybrid female sterility. Molecular ecology, 27(4), 949-958.

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