mucispirillum schaedleri BioVector NTCC质粒载体菌种细胞基因保藏中心

mucispirillum schaedleri BioVector NTCC质粒载体菌种细胞基因保藏中心
Designation: HRI I17 [ACM 5223]
Medium
Medium 260: Trypticase soy agar/broth with defibrinated sheep blood
Growth Conditions
Temperature: 37°C
Atmosphere: Anaerobic; moist conditions
Propagation Procedure
1. This organism is shipped frozen in dry ice. Just prior to use, thaw vial in water at approximately 37oC.
When thawed, a drop of the suspension may be used to do an immediate wet mount to observe the unique
morphology of this organism and verify its viability by checking for motility.
2. Aseptically transfer the thawed suspension into a fresh #18 broth (35
ml). Mix well. This suspension can
now be used to inoculate agar slant(s), plate(s), or the preferred biphasic culture. Two #260 plates should
be inoculated, one for anaerobic growth and the second for aerobic growth. No growth should occur on the
plate incubated aerobically.
3. To obtain a biphasic culture, add 0.6 ml of the suspension to a # 260 slant. The resulting pool at the bottom
of the slant is where the best, most rapid growth will occur.
4. Incubate the broth tubes, slants, and plates at 37oC under anaerobic conditions. You may use an anaerobe
jar with catalyst and gas generator pack or other suitable means of producing anaerobic conditions. Be sure
caps are loosen to facilitate gas exchange. Incubate aerobic blood plate aerobically at 37oC.
5. Within 35
days, good growth should be obtained in the broth pool at the bottom of the slant. Additional
incubation may be required for colonies to appear on agar plate. Further subcultures can be made using the
broth pool as the inoculum source. Subcultures to biphasic cultures will require only 24 to 48 hours of
incubation for good growth.
Always use freshly prepared prereduced
media or prereduced
media that has been previously
prepared but stored under anaerobic conditions.
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