| Autor: |
Jeannottee, Richard, Lee, Eric, Arabyan, Narine, Kong, Nguyet, Kao Thao, Huang, Carol, Storey, Dylan, Weimer, Bart, Kelly, Lenore |
| Rok vydání: |
2015 |
| DOI: |
10.6084/m9.figshare.1372500.v1 |
| Popis: |
Shearing of bacterial gDNA within a specific size range prior to sequencing libraryconstruction is a critical step in Next Generation Sequencing workflows. The qualitycontrol of the sheared bacterial gDNA is required in large multiplexed formats forlarge volume workflows, such as those used in the 100K Pathogen GenomeSequencing Project. Using the Covaris E220 instrument, the power and treatmenttime were varied to determine the effect on the optimal fragment size (150–350 bp)in the resulting sheared gDNA of four bacterial pathogens: Salmonella entericasubsp. enterica serovar Saint Paul strain Sp3 and serovar Typhimurium strain LT2,Klebsiella sp. and Vibrio spp. DNA fragment quantification and sizing were mea-sured using an Agilent 2200 TapeStation system, and Agilent High Sensitivity D1000ScreenTape assay. The 2200 TapeStation system was suitable to determine size dis-tribution after fragmentation of gDNA in a 96-well plate format, a format suitable for high-throughput workflow and compatible with shearing technologies that use a 96-well plate multiplexed format. This approach enabled the measurement of gDNAand sheared DNA using a single technology. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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