| Autor: |
Xu J; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Pandoh PK; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Corbett RD; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Smailus D; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Bowlby R; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Brooks D; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., McDonald H; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Haile S; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Chahal S; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Bilobram S; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Mungall KL; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Mungall AJ; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Coope R; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Moore RA; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Zhao Y; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Jones SJ; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada., Marra MA; Canada's Michael Smith Genome Sciences Centre at BC Cancer, 570 W 7th Ave, Vancouver, Canada.; Department of Medical Genetics, University of British Columbia, 2329 West Mall, Vancouver, Canada. |
| Abstrakt: |
High-throughput total nucleic acid (TNA) purification methods based on solid-phase reversible immobilization (SPRI) beads produce TNA suitable for both genomic and transcriptomic applications. Even so, small RNA species, including miRNA, bind weakly to SPRI beads under standard TNA purification conditions, necessitating a separate workflow using column-based methods that are difficult to automate. Here, an SPRI-based high-throughput TNA purification protocol that recovers DNA, RNA and small RNA, called GSC-modified RLT+ Aline bead-based protocol (GRAB-ALL), which incorporates modifications to enhance small RNA recovery is presented. GRAB-ALL was benchmarked against existing nucleic acid purification workflows and GRAB-ALL efficiently purifies TNA, including small RNA, for next-generation sequencing applications in a plate-based format suitable for automated high-throughput sample preparation. |
