Experiment 1: Preparation of MS (Murashige and Skoog) Media
Aim:
To prepare MS medium used for plant tissue culture.
Requirements:
MS salts, sucrose, vitamins, plant growth regulators (e.g., auxins and cytokinins), distilled
water, pH meter, NaOH/HCl, autoclave, agar (for solid medium).
Procedure:
1. Weigh and dissolve required quantity of MS salts in ~800 ml of distilled water.
2. Add vitamins and 3% sucrose (w/v) to the solution.
3. Add plant growth regulators as needed depending on the plant species and purpose.
4. Adjust the pH of the medium to 5.8 using 0.1N NaOH or HCl.
5. Add 0.8% agar if a solid medium is desired.
6. Make up the final volume to 1000 ml with distilled water.
7. Dispense the medium into culture vessels.
8. Sterilize by autoclaving at 121°C and 15 psi for 15–20 minutes.
Result:
MS medium was successfully prepared and sterilized for use in plant tissue culture.
Precautions:
Handle all chemicals with care and wear protective gear.
Maintain aseptic conditions to avoid contamination.
Ensure correct pH adjustment for optimal plant growth.
Autoclave properly to sterilize the medium.
Experiment 4: Micropropagation of Potato (Solanum tuberosum)
Aim:
To perform micropropagation of potato using in vitro techniques.
Requirements:
Sterilized explants (e.g., shoot tips or nodes), MS medium, laminar airflow cabinet,
sterilization tools, culture tubes, autoclave.
Procedure:
1. Select healthy potato plants and excise shoot tip or nodal explants.
2. Surface sterilize explants using 70% ethanol for 30 seconds followed by 0.1% HgCl₂ for
2–3 minutes.
3. Rinse explants 3–4 times with sterile distilled water.
4. Inoculate explants on MS medium supplemented with appropriate growth regulators
(e.g., BAP).
5. Incubate cultures at 25±2°C with a 16-hour light/8-hour dark photoperiod.
6. Observe shoot proliferation and subculture after 2–3 weeks for further multiplication.
Result:
Potato explants showed shoot proliferation indicating successful micropropagation.
Precautions:
Use sterile instruments and environment to prevent contamination.
Do not overexpose explants to sterilizing agents.
Maintain consistent temperature and light conditions in the culture room.
Label all culture tubes properly.
Experiment 7: To Find Out Specific Gravity of Soil
Aim:
To determine the specific gravity of a soil sample.
Requirements:
Pycnometer, balance, distilled water, dry soil sample.
Procedure:
1. Clean and dry the pycnometer and weigh it (W1).
2. Fill it with dry soil and weigh (W2).
3. Add distilled water to the pycnometer containing soil and weigh again (W3).
4. Fill the pycnometer with only distilled water and weigh (W4).
Result:
Specific gravity of the soil sample was calculated accurately using the pycnometer method.
Precautions:
Ensure the pycnometer is clean and dry before use.
Avoid air bubbles while filling with water.
Use accurate weighing techniques.
Repeat readings for more accurate results.