A Simple Preparation of Mycoplasmal DNA Template for PCR from Biological Samples Using Effective Surfactants.

An improved loop-mediated isothermal amplification assay for the detection of <i>Mycoplasma bovis</i>

“…Mycoplasmas were cultured in M broth [ 10 ] at 37°C in a 5% CO 2 environment. After incubation, cells per 500 µl of culture medium were pelleted, resuspended in 25 µl of mycoplasma lysis buffer containing proteinase K [ 11 ], and incubated at 60°C for 50 min (for DNA extraction) and then at 100°C for 10 min (for proteinase K inactivation). The non-mycoplasma strains were grown overnight on Mueller-Hinton Agar (MHA, OXOID, Hampshire, U.K.) plates, and single colonies of each strain were employed for DNA extraction.…”

Detection of Mycoplasma in Mastitic Milk by PCR Analysis and Culture Method.

“…The pellet was resuspended with 1.0 ml of PBS and centrifuged at 6,000 rpm for 10 min. The pellet was resuspended with a 50 ml aliquot of mycoplasmal lysis buffer [14] containing 200 µg/ml of proteinase K, incubated at 60°C for 1 hr, heated at 100°C for 5 min to inactivate the proteinase K, rapidly chilled on ice, and stored at -20°C until use. This sample was also used as a template DNA on PCR.…”