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BKMAM can accept OEM and ODM orders.

SOLUTIONS 1 CELL CULTURE-PRIMARY/SUBCULTURE
SOLUTIONS 2 MICROBIAL CULTURE PROCESS
SOLUTIONS 3 MOLECULAR DETECTION-WESTERN BLOT EXPERIMENT

// SOLUTIONS 1 CELL CULTURE-PRIMARY/SUBCULTURE

01 Culture - cut tissue

- Cut the tissue into small pieces under aseptic conditions.
- Place them in a culture dish or a small beaker.
- Wash twice with PBS buffer to remove blood cells and debris.

02 Culture - digestion separation

- Add the tissue and trypsin to cutting tube, and digest in 37°C water bath. During digestion, pipette periodically to promote digestion.
- After digestion is complete, add complete medium/medium + fetal bovine serum + double antibody to terminate the digestion. Filter the cell suspension and collect the fluid.

03 Culture - centrifugation

- Centrifuge the filtrate, discard the supernatant, and retain the double antibody to terminate the digestion. Filter the cell suspension and collect the fluid.

04 Primary culture - culture

- Resuspend the cells in complete medium (blood medium + serum + double antibody) and transfer them to a culture flask.
- Incubate in 37°C, 5% CO2 incubator.

05 Subculture - subculture medium change

- For subculture of primary cells, discard the medium, rinse with PBS, add in appropriate amount of trypsin/glucose solution, and digest at 37°C for 1-2 minutes.
- Wash cells between rounds of incubation and remove with a pipette, immediately add complete medium to terminate the digestion. Perform trypsin in an overnight suspension. After centrifugation, resuspend the cells, perform counting, dilute at an appropriate ratio, and resuspend in complete medium.

// SOLUTIONS 1.1 CELL CULTURE-CELL CRYOPRESERVATION AND THAWING

01 Preparation and pre-freezing solutions

Select an appropriate cryopreservation solution (left-cup/preserved formula medium, serum, DM50-7-7-2-1, or a commercial cell cryopreservation solution).

02 Resuspending and aliquoting

- Liquefy or isolate cells in the appropriate growth media, after centrifugation, completely dissolve the supernatant. 2. Centrifuge the sample to pellet in cryopreservation solution, adjust to high concentration, aliquot into cryopreservation tubes.

03 Programmed cryopreservation

- Place the cryovials into a programmed cooling container and keep at -80°C overnight, then transfer to liquid nitrogen for long-term storage.

04 Rapid thawing

Remove the cryovials from liquid nitrogen, warm overnight, and immediately thaw in a 37°C water bath. Gently shake for no more than 1 minute until completely thawed.

05 Inoculation culture

- Transfer the cells suspension to a new 96-well culture chamber using a new volume of complete medium, new well, or fresh, and incubate for 24 hours. 2. Resuspend the cells in fresh medium and seed into a new culture flask. It is recommended to change the medium 24 hours after recovery.

// SOLUTIONS 1.2 CELL CULTURE-CELL COUNTING

01 Cleaning the counting plate

- Use less cleaning paper or fine paper to reduce the amount of waste.
- Use a clean and dry bench to avoid cross-contamination.
- Use a disposable cutting mat to avoid cross-contamination.
- Use a disposable cutting mat to avoid cross-contamination.

02 Suspension preparation and staining

- Use the suspension solution with a suitable volume of 0.5 mL.
- Add the suspension solution to the sample plate.
- Add the sample solution to the sample plate.

03 Filler cell counting chamber

- Place the cryovials into- After mixing the stained cell suspension, slowly add a large volume of the cell suspension.
- Add the cell suspension to the sample plate.
- Add the sample solution to the sample plate. a programmed cooling container and keep at -80°C overnight, then transfer to liquid nitrogen for long-term storage.

04 Microscopic counting

- Place the counting chamber under a microscope and count the cells within the four corners of the sample plate.
- Use a microscope to count the cells within the four corners of the sample plate.
- Use a microscope to count the cells within the four corners of the sample plate.
- Use a microscope to count the cells within the four corners of the sample plate.

// SOLUTIONS 2 MICROBIAL CULTURE PROCESS

01 Sample pretreatment

- Sample was collected using sterile tools, solid samples were homogenized, and then acidified by adding 6N HCl.

02 Preparation and sterilization

- Washing and crushing: Wash the sample with water according to the culture medium formula and add distilled water to a beaker.
- Disinfecting: Disinfect the sample with 0.1% sodium hypochlorite solution for 30 minutes.
- Sterilization: Sterilize the disinfected culture medium in an autoclave at high temperature and pressure. After sterilization, store or perform subsequent operations such as plate testing.

03 Microbial inoculation

- Using sterile conditions, the target microorganism is inoculated onto the surface of a solid culture medium or into a liquid culture medium.

04 Microbial culture

- Place the inoculated culture medium in a constant temperature incubator, and a suitable temperature, and the incubation time depends on the type of microorganism.

05 Observation and recording of culture results and preservation of microbial strains

- Observe and record the growth of the microorganism, and preserve the results.

// SOLUTIONS 3 MOLECULAR DETECTION-WESTERN BLOT EXPERIMENT

01 Sample Preparation

- Protein extraction: Wash the sample with PBS buffer, add an appropriate amount of Dolly buffer for Dolly, and then add the sample to the membrane.
- Protein quantification: Determine the protein concentration of the total protein solution using a BCA protein quantification kit.

02 Gel Electrophoresis

- Add the sample and protein marker into the prepared gel loading wells, and start the power supply for electrophoresis separation.

03 Membrane Transfer

- Separate the proteins on the gel and transfer them in situ onto a solid support membrane. Place the prepared plate into the transfer apparatus. After transfer, wash the membrane.

04 Closed

- Place the transferred membrane into an incubation box, add 5% non-fat milk solution or BSA blocking buffer, and incubate for 1 hour.

05 Antibody Incubation

- Place the blocked membrane into the primary antibody solution for incubation. After incubation, wash the membrane, then transfer it to the secondary antibody solution for incubation. Following this, wash the membrane again.

06 Detecting Development

- Depending on the type of chromosome or Univesece detection, select an appropriate gel imaging system or bioluminescence imaging instrument for visualization.

BKMAM can accept OEM and ODM orders.

// SOLUTIONS 1 CELL CULTURE-PRIMARY/SUBCULTURE

// SOLUTIONS 1.1 CELL CULTURE-CELL CRYOPRESERVATION AND THAWING

// SOLUTIONS 1.2 CELL CULTURE-CELL COUNTING

// SOLUTIONS 2 MICROBIAL CULTURE PROCESS

// SOLUTIONS 3 MOLECULAR DETECTION-WESTERN BLOT EXPERIMENT

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