We can all be scientists given the right equipments, facility and instructions. All the bogus terminologies you hear and get scared are really all basic things that are around you everyday. Today, I'll be taking you with me to the lab to show you how I extract DNA, be sure to don your PPE and enjoy the lab.

Here's a brief introduction so you can know what you are doing.

DNA EXTRACTION

DNA isolation is a process of purification of DNA from sample using a combination of physical and chemical methods. The first isolation of DNA was done in 1869 by Friedrich Miescher.

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DNA EXTRACTION

DNA is extracted from human cells for a wide range of reasons. A pure sample of DNA enables you to test for a genetic disease in a new born, analyze forensic evidence, and or study a gene involved in cancer.

Scientists can buy ready-to-use DNA extraction kits. These kits help extract DNA from particular cell types or sample types. Being able to extract deoxyribonucleic acid (DNA) is important for a number of reasons. By studying nucleic acid, we can identify genetic disorders or diseases, and also possibly find cures for them by manipulating or experimenting with nucleic acid.

There are several methods employed in the extraction DNA.
DNA Extraction Methods from Blood Impregnated Filter Paper; Methanol-Heat Extraction Method.

Equipments and Materials:

• Double distilled water/nuclease free water.
  Absolute methanol.
• Heat block.
• Vortex.
• Cold centrifuge.
• 1.5 ml microfuge tubes.
• Pipette tips and pipette of various calibration.
• Samples.

Protocol:

• A portion of blood impregnated filter paper was cut into 1.5 ml microfuge tube. The filter paper was chopped into small bits.
• 100 µl of absolute methanol was added.
• It was allowed to fix/inactivate for 20 minutes.
• The methanol from the tube was discarded taking caution to avoid losing the filter papers.
• The filter paper was allowed to dry properly.
  100 µl of double distilled water was added.
• It was heated at 94oC for 20 minutes, vortexing every 3 minutes.
• It was centrifuged for 2 minutes.
• 5 µl is used for PCR.
• The remaining was stored at -20°C till required.

DNA Extraction Methods from Blood Impregnated Filter Paper; Qiagen Extraction Kit Method.

Equipments and Materials:

• Ethanol.
• Water bath.
• Vortex.
  
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• Cold centrifuge.
• 1.5 ml microfuge tubes.
• Pipette tips and pipette of various calibrations.
• Qiagen spin column.
• Samples.

Reagents:

• Qiagen protease stock solution.
• Dissolve the lyophilized Qiagen protease with 1.2 ml protease solvent (store at 2-8oc or -20oc).
• Buffer AL (lysing).
• Buffer AW1 (washing). Add 25 ml Ethanol to the 19 ml Buffer AW1 concentrate to give a final volume of 44 ml.
• Buffer AW2 (washing). Add 30 ml of Ethanol to the 13 ml of Buffer AW2 concentrate to give a final volume of 43 ml.
• Buffer AE (elution).

Procedure:

• I cut dried blood spot into a 1.5 ml microcentrifuge tube.
• I then added 180 µl of Buffer AL.
• It was incubated at 85°c (water bath) for 10 minutes and centrifuged briefly.
• 20 µl of proteinase K was added, it was then mixed and incubated at 56oc for 1 hour. I then centrifuged briefly.
• I added 200 µl Buffer AL, mixed and incubated at 70oc for 10 minutes. I then centrifuged briefly.
• I added 200 µl ethanol to the sample, mixed and centrifuged briefly.
• I transfered the content into the spin column.
  I centrifuged at 6, 000xg for 1 minute.
• I placed the spin column into a new collection tube and discarded the old collection tube with the content.
• I added 500 µl Buffer AW1 and centrifuged at 6, 000xg.
• I repeated step 9.
• I added 500 µl Buffer AW2 and centrifuged at 14, 000xg for 3 minutes.
• I placed the spin column into a new collection tube and discarded the old collection tube with the content.
• I added 100 µl Buffer AE.
• I incubated at room temperature for 1 minute.
• I centrifuged at 8, 000xg for 1 minute.
• 5 µl was used as DNA template for PCR and the remaining stored at -20°c.

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