Praktikum Genetika Ternak - Ekstraksi DNA Darah

Introduction

This tutorial provides a step-by-step guide on how to extract DNA from blood samples, as demonstrated in the practical genetics workshop for livestock. Understanding DNA extraction is crucial for various genetic analyses, including genetic markers, individual identification, and DNA-based selection in animal breeding.

Step 1: Gather Necessary Materials

Before starting the DNA extraction process, ensure you have the following materials ready:

• Blood sample (preferably fresh)
• Sterile tubes or containers for sample storage
• Lysis buffer (to break open cells)
• Proteinase K (to digest proteins)
• Ethanol or isopropanol (for DNA precipitation)
• TE buffer or distilled water (for DNA resuspension)
• Pipettes and tips
• Centrifuge
• Vortex mixer
• Microcentrifuge tubes

Step 2: Prepare the Blood Sample

• Place the blood sample in a sterile tube.
• If the sample is not fresh, store it at appropriate temperatures to maintain DNA integrity.
• Mix gently to avoid cell rupture.

Step 3: Cell Lysis

• Add an appropriate volume of lysis buffer to the blood sample. This buffer contains detergents that will break down cell membranes.
• Add Proteinase K to the mixture. This enzyme will help digest proteins and facilitate DNA extraction.
• Incubate the mixture at 56°C for 1-2 hours to ensure complete lysis of the cells.

Step 4: DNA Precipitation

• After incubation, cool the sample to room temperature.
• Add an equal volume of ethanol or isopropanol to the lysate. This step precipitates the DNA.
• Mix gently by inverting the tube several times or using a vortex mixer to ensure thorough mixing.

Step 5: Centrifuge the Sample

• Centrifuge the tube at high speed (around 12,000-15,000 rpm) for 10-15 minutes.
• This process will separate the DNA from the solution, forming a pellet at the bottom of the tube.

Step 6: Remove Supernatant

• Carefully pour off the supernatant without disturbing the DNA pellet.
• Wash the pellet with 70% ethanol to remove any remaining impurities.
• Centrifuge again for a few minutes and discard the ethanol.

Step 7: Resuspend DNA

• Allow the DNA pellet to dry briefly.
• Resuspend the pellet in TE buffer or distilled water.
• Store the DNA at -20°C for long-term storage.

Conclusion

You have successfully extracted DNA from a blood sample! This process is fundamental in genetics studies involving livestock. Key takeaways include the importance of using proper materials and maintaining the integrity of your samples throughout the procedure. For further applications, consider exploring genetic marker analysis or individual identification in your research. Happy experimenting!