DIVA DNA Sequence Validation

DIVA DNA Sequence Validation

 

Overview: 

Comprises sequence validation of plasmid and/or amplicon DNA using the Illumina MiSeq platform. Customized ABF workflow minimizes library preparation steps and leverages laboratory automation to enable scale/reduce reagent costs, provides full plasmid coverage, and is integrated with the ICE sequence/strain repository platform. This validates DNA sequences (ensures that they actually are what they purport to be) at effective cost points and turnaround times and prevents incorrect materials from being wastefully investigated or processed further. The process consists of weekly cycles of 384 samples (individually barcoded and then pooled) each. Samples can consist of purified plasmids, PCR amplicons, or (plasmids in) bacterial cultures. There are some constraints around acceptable resuspension buffers, minimum concentrations, minimum volumes, etc. Available internally to ABF researchers, as well as to ABF CRADA projects. 

References and Additional Information: 

Thompson et al. (2018) “Isolation and characterization of novel mutations in the pSC101 origin that increase copy number”. Scientific Reports 8, 1590 doi:10.1038/s41598-018-20016-w

Labs:

Lawrence Berkeley National Laboratory

Categories: Build