Tweet
Recently Ilumina released additional sets of barcoded Nextera primers which enable multiplexing 384 samples for sequencing. High level of multiplexing for sequencing on NextSeq 500 system High output kit sometimes is essential as lanes can not be clustered separately. Equimolar pooling of that many libraries by current gold standard (QPCR+ average library sizing by Bioanalyzer or TapeStation) is a daunting task. A possible work around to avoid hundreds of QPCR is to use beads for normalisation when pooling libraries of similar size distribution in which the equal mass (volume) pooling is equimolar pooling followed by QPCR quantification of pool. Currently there are two products for normalisation. They work by binding (switching charge by pH) or precipitating DNA (PEG+salt) onto beads with low capacity for interaction with DNA. AMPure beads have high capacity and cannot be saturated by concentrations of most NGS libraries. It will be good if anyone with experience using this normalisation products or homebrew versions to share their experience with Seqanswers community.
-
Last edited by nucacidhunter; 05-08-2014, 06:24 AM.
-
Cancer research has been transformed through numerous molecular techniques, with RNA sequencing (RNA-seq) playing a crucial role in understanding the complexity of the disease. Maša Ivin, Ph.D., Scientific Writer at Lexogen, and Yvonne Goepel Ph.D., Product Manager at Lexogen, remarked that “The high-throughput nature of RNA-seq allows for rapid profiling and deep exploration of the transcriptome.” They emphasized its indispensable role in cancer research, aiding in biomarker...09-07-2023, 11:15 PM -
Ribonucleic acid (RNA) represents a range of diverse molecules that play a crucial role in many cellular processes. From serving as a protein template to regulating genes, the complex processes involving RNA make it a focal point of study for many scientists. This article will spotlight various methods scientists have developed to investigate different RNA subtypes and the broader transcriptome.
Whole Transcriptome RNA-seq
Whole transcriptome sequencing...08-31-2023, 11:07 AM