Streptococcus pneumoniae|BNCC338425 |BNCC

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Streptococcus pneumoniae

Culture medium	Columbia blood plate (CA-B): casein pancreatin digest 10.0g, cardiopancreatin digest 3.0g, corn starch 1.0g, meat gastric enzyme digest 5.0g, yeast extract 5.0g, sodium chloride 5.0, agar 20.0g, distilled water 1.0L,pH 7.3±0.2. After sterilization at 121 ℃ for 15min, cool to 55 ℃, add 5% sterile defibrated sheep blood, shake well, pour the plate and cool it for later use.
Subculture procedure	(1) Prepare 1-2 pieces of the plate; (2) sterilize the surface of the plate and open it in biosafety cabinet; (3) Absorb 1-3ml of sterile water into the plate, scrape the bacterial lawn back and forth on the surface of the plate with a sterile spreader to make a bacterial suspension; (4) suck the bacteria suspension with a pasteur pipette, dispense it into a fresh plate evenly. (5) put the plates under the growth conditions.
Growth conditions	37 ℃;24-48 hours; 5% CO2
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, opaque, golden yellow on the front and golden yellow on the back, flat shape, smooth and bright surface, moist texture, easy to stir up, G + (blue-purple), cocci, purity: pure
application	Research
Sharing mode	Public welfare sharing

Streptococcus pneumoniae

SStorage conditions: 2~8 ℃

No. 338425

Product format: freeze dried, 200ul

Validity period: 6 years

Biosafety level: 2 , handle in safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the cells are well. For the frozen cells, it shall be stored in the refrigerator at -80 ℃ upon arrival. If they are not used for a long time, they shall be transferred to liquid nitrogen for storage overnight. Recovered cells in T25 culture flask, upon receipt, put the culture flask in the incubator for 4h, and then carry out handling procedure. During recovery, each vial shall be used up once and shall not be retained. After recovery, the cells can be passed on to the next generation and can be used normally. Please operate in strict accordance with this instruction, otherwise the replacement of cells are not be available in case of lose of cell viability.

Growth conditions:37 ℃,5% CO₂, Columbia blood plate (ready-to-use type),24-48h.

Recovery steps:

(1)Prepare 1-2 of above mentioned plates;

(2)Open the biosafety cabinet, sterilize of the ampoule, heat the tip in a flame, quickly drop sterile water to creak it, then break it with forceps;

(3)Draw 0.5ml of sterile water into the freeze dried ampoule, make it fully dissolved, and distribute the solution to the plates well in 200ul/plate;

(4)Put the plates under the above culture conditions for cultivation for 24-48 hours.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

item	test result
viability:	good viability, in 24-48h liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology: (above)	size: 1-2mm shape: round edge: neat transparency: opaque color: off-white uplift: middle raised surface: bright and smooth texture: moist and viscous
Conclusion:	good viability, no abnormal colony morphology, qualified

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