ISOLASI DNA Praktik Laboratorium Biologi Molekuler -BAGIAN 1

Summary

This video demonstrates the process of DNA isolation from blood. It involves taking 2-3 mL of blood, mixing it with RBCLB, and homogenizing before placing it on ice for 20 minutes. The sample is then centrifuged at 5,000 rpm for 10 minutes, with the supernatant discarded. After cleaning the tube, the pellet is re-suspended in 100 µL of SE buffer and transferred to a 1.5 mL Eppendorf tube for further processing. This procedure ensures the extraction of clean DNA for downstream applications.

Takeaways

• 🧬 The procedure focuses on DNA isolation from a blood sample.
• 🧪 Begin by taking 2–3 mL of blood and placing it in a Falcon tube.
• 💧 Add 8–12 mL of RBCLB (Red Blood Cell Lysis Buffer) to the blood sample.
• 🥶 Homogenize the mixture thoroughly and then keep it on ice for 20 minutes to maintain stability.
• ⚙️ Centrifuge the mixture at 5,000 rpm for 10 minutes to separate components.
• 🧻 After centrifugation, discard the supernatant carefully to retain the pellet containing the desired material.
• 🧼 Clean the inner wall of the tube using a tissue to remove residual liquid.
• 💦 Add 100 µL of SE buffer to the pellet to dissolve it completely.
• 🔄 Mix or shake the tube until the pellet is fully dissolved in the SE buffer.
• 📦 Transfer the dissolved solution into a 1.5 mL Eppendorf tube for further DNA processing or storage.

Q & A

What is the main purpose of the procedure described in the transcript?

The main purpose of the procedure is to isolate DNA from a blood sample using a series of preparation and centrifugation steps.

How much blood is required for the DNA isolation process?

Approximately 2 to 3 milliliters of blood are needed for the process.

What type of tube is initially used to collect the blood sample?

A Falcon tube is used to collect and process the blood sample initially.

What is added to the blood sample after collection, and why?

8 to 12 milliliters of RBCLB (Red Blood Cell Lysis Buffer) are added to lyse red blood cells and release cellular components for DNA isolation.

Why is the mixture placed on ice for 20 minutes?

The mixture is placed on ice to stabilize the sample and slow down enzymatic degradation, ensuring better DNA preservation.

At what speed and duration is the centrifugation step performed?

The sample is centrifuged at 5,000 revolutions per minute (rpm) for 10 minutes.

What should be done with the supernatant after centrifugation?

The supernatant should be discarded carefully, leaving only the pellet that contains the desired cellular material for further processing.

How is the tube cleaned after removing the supernatant?

The walls of the tube are cleaned by gently turning the tube onto a tissue to remove any residual liquid without disturbing the pellet.

What is the purpose of adding 100 µL of SE buffer to the pellet?

SE buffer is added to dissolve and resuspend the pellet, preparing it for further DNA purification steps.

Where is the dissolved solution transferred after resuspension?

The resuspended solution is transferred into a 1.5 mL Eppendorf tube for the next stages of DNA isolation and analysis.

What precautions should be taken during the DNA isolation procedure?

Samples should be kept on ice when required, handled gently to avoid DNA shearing, and processed using sterile equipment to prevent contamination.