I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the positive controls in the Bacterial 500 and the Fungal kits have worked satisfactorily in the assay, so the PCR reaction, cycle sequencing reaction and associated clean up reactions seem to be working ok. Do you have any recommendations for suitable genomic DNA extraction protocols that work well for MicroSeq ID? Is it likely that the standard Prepman Ultra extraction protocol I am using is either not efficient or not producing a suitably good quality gDNA template for PCR? Please let me know if you require further information.
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