Space enterococcus faecium LCT-EF258 strain
A technology of LCT-EF258 and Enterococcus faecium, which is applied to the determination/testing of bacteria and microorganisms, microorganisms, etc., and can solve problems such as the lack of a good research entry point
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Embodiment 1
[0029] Implementation example one: Gram staining of Enterococcus faecium LCT-EF258 bacterial strain
[0030] Take 50ul of the sample and centrifuge, discard the supernatant, add 50ul of sterile water, absorb 20ul and fix it on the glass slide; for initial staining, absorb 2 drops of Gram stain I (crystal violet) on the glass slide sample for initial staining for 1min, Then rinse the dye solution with tap water; for mordant dyeing, absorb 2 drops of Gram stain II (iodine solution) to cover the painted surface and dye for about 1min, then rinse the dye solution with tap water, and absorb the water on the coated surface with absorbent paper; for decolorization, absorb Gram Put 2 drops of Gram's stain III (alcohol) on the coated surface for about 30 seconds, rinse the glass slide with water, and absorb the water on the coated surface with absorbent paper; for counterstaining, absorb 2 drops of Gram stain IV (safranin) After about 1 minute on the surface, rinse the glass slide with...
Embodiment 2
[0031] Implementation example two: Antibiotic susceptibility detection of Enterococcus faecium LCT-EF258 strain
[0032] Prepare 25L of LB solid medium, pour 1500 plates after sterilization, dry, and set aside; for the preparation of bacterial suspension, take out the slant of these single bacteria, and prepare a 1.5ml centrifuge tube filled with 1ml sterile saline, and use Dip a sterilized bamboo stick into a 1.5ml centrifuge tube, mix well, and adjust the bacterial concentration to about 10 7 ~10 8 ;Dilution and coating, draw 100 μl of the prepared bacterial suspension for coating, spread as evenly as possible, apply 6 flat plates for each single strain of bacteria, and proceed to the next step (paste the drug-sensitive sheet) after each strain of bacteria is coated ;Paste the drug-sensitive sheet, select antibiotics for drug-sensitivity test to screen out the strains with drug-resistant mutations; after the drug-sensitive sheet is pasted, place the plate at 37°C for 18-24 ...
Embodiment 3
[0033] Implementation Example 3: Physiological and biochemical identification experiment of Enterococcus faecium LCT-EF258 strain
[0034] To prepare the bacterial suspension, take out the slant of the strain, use a sterilized bamboo stick to pick up the strain and inoculate it in LB liquid culture medium (4ml system), and cultivate for 24h. Then take 2ml of the bacterial solution and centrifuge at 6000r / s for 5min, discard the supernatant medium, leave the centrifuged bacteria, then wash the bacteria with sterilized normal saline, centrifuge at 6000r / s for 5min, discard the supernatant again, and use Biolog The inoculum was used to suspend the bacteria, and finally transferred to 15ml of the inoculum. Adjust to a concentration of about 10 7 ~10 8 ; Inoculate, pour the above bacterial suspension into the inoculation dish, shake evenly, add samples with 12 (or 8) row guns, add 100ul to each Biolog plate hole, a total of 96 holes, after adding the sample, cover Cover the Biol...
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