These protocols describe methods to extract high molecular weight genomic DNA from Bacillus subtilis (strain CECT® 356), as an example of gram-positive bacteria. We recommend QIAGEN Genomic-tip 500/G, QIAGEN MagAttract HMW DNA Kit, and QIAGEN Puregene Cell Kit. The Genomic-tip generated high sequencing output and long read lengths. For users seeking a simple workflow, we obtained high sample recovery and high sequencing output with the MagAttract HMW DNA Kit, and while longer reads were obtained with the QIAGEN Puregene Cell Kit, sample recovery was much reduced. We also recommend the use of SPRI size selection prior to sequencing. The sequencing performance was assessed using the Ligation Sequencing Kit.

Please see below a comparison of DNA fragment lengths, sequencing read lengths, and throughput for the three protocols. Click on the panel of your choice to view the protocol.