Crude toxin supernatants with equivalent toxin protein amounts are listed. Values included in this table are the exact copy number of BoNT DNA detected in crude toxin preparations at the indicated amounts of protein. LOD indicates the averaged limits of detection for that subtype in our mouse protection bioassay with identical serotypes used in toxin complex preparations. Next, we did comparative testing of crude culture supernatants

(without DNA extraction) against purified DNA preparations from the same strains. As the crude culture supernatants contained botulinum selleck chemicals llc neurotoxins, they were tested at an independent location DAPT molecular weight that is registered for the use of botulinum neurotoxins using alternative equipment (the Roche Light Cycler versus the ABI 7700 for the purified DNA preparations). All 23 BoNT-containing samples tested positive for the appropriate toxin subtype, including three samples containing multiple toxin serotypes (A2b, Ba4, and Bf). In addition, the mosaic C/D and D/C strains had positive PCR signals for both serotype C and D, confirming the existence of both BoNT/C and/D gene sequences in these strains. The results, shown in Table 6, indicate that this assay is equally effective at detecting and identifying BoNT genes regardless of the sample (crude culture supernatants

or purified DNA preparations) or the equipment used. Table 6 Detection of BoNT DNA from purified DNA of bacterial cultures or extracted DNA from crude toxin supernatants     BoNT A BoNT B BoNT C BoNT many D BoNT E BoNT F BoNT G BoNT subtype strain ABI LC ABI LC ABI LC ABI LC ABI LC ABI LC ABI LC A1 Hall ++++ +++                         A2b CDC 1436 ++ ++++   +++                     A3 Loch Maree ++ ++++                         B1 Okra     ++++

+++                     B2 213B     ++++ ++                     B2 CDC 1828     ++++ +++                     B3 CDC 795     +++ +++                     B4 (npB) Eklund 17B     ++ +++                     Ba4 CDC 657 + + +++ +++                     Bf An436     +++ +++             ++ +++     C Stockholm         ++++ +++                 C/D 6813         ++ ++ ++               D ATCC 11873             ++ +++             D/C VPI 5995         ++   ++++ +++             E1 Beluga                 ++++ ++         E2 CDC 5247                 ++++ ++         E2 CDC 5906                 +++ ++         E3 Alaska E43                 ++++ +++         E4 (It butyr) BL5262                 +++ ++         F1 (prot) Langeland                     ++++ +++     F2 (np) Eklund 202F                     +++ ++     F3 (baratii) Orange                     ++       G 1354                         ++++ +++ C.