Introduction of Blood and Tissue DNA Extraction
Table of Contents
Advances in DNA technology have been created in the increased application of DNA tests. From paternity determination to criminal investigations and even disease diagnosis the accuracy and data provided mean that millions of people are now shifting to DNA testing to solve their questions.
Test Requirements for Blood and Tissue DNA Extraction
DNA extraction kit and it contains-
- Proteinase K
- Lysis buffer (AL)
- Wash solution (AW1)
- Wash solution 2 (AW2)
- Elution buffer(AE)
- Spin column and collection tubes
Extra we need-
- Ethanol (molecular grade)
- Centrifuge tubes
- Centrifuge
- Short spinner
- Micropipettes and tips
- Waste bin
- Tissue paper roll
DNA Extraction Procedure
- Place 20 μL proteinase k in an Eppendorf tube of 1.5 ml and then put a 200 ul sample (blood).
- Add 200 μL AL buffer and vortex for 15 seconds and incubate at 56 C for 10 minutes.
- Perform a short spin.
- Add 200 μL absolute ethanol of molecular grade and again vortex or short spin for 15 seconds.
- Transfer the mixer to a spin column and centrifuge at 8000 rpm/ 6000g for a minute.
- Transfer the column to the collection tube provided 500 ul AW1 and again centrifuged at 8000 rpm or 6000g for a minute.
- Discard the collection tube.
- Add 500 μL of AW2 and centrifuge at 14000 rpm for 3 minutes.
- Discard the collection tube.
- Transfer to a new 1.5 ml centrifuge tube.
- Add 200 μL AE buffer and centrifuge at 8000 rpm for a minute.
- This is a solution having DNA i.e. template.
Keynotes
- The template should be used on the same day otherwise it should be stored as genomic DNA stored at -20°C and -80°C is of good quality, and these specimens withstand multiple freeze-thaw cycles.
- DNA quality can be assessed by agarose gel electrophoresis, PCR amplification of an
the indicator housekeeping gene (β-globin), and SNP assays on various platforms. - For short-term studies, genomic DNA can be stored at 4°C or even room temperature (RT) without degradation, but specimens should be monitored for DNA concentration and evaporation.
- DNA Extractor Automations are also available in the market to minimize the cumbersome steps of manual extraction.
Further Readings
- https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=&cad=rja&uact=8&ved=2ahUKEwiD_f2AjZP1AhUu7XMBHUpMAzQQFnoECAQQAQ&url=https%3A%2F%2Fwww.qiagen.com%2Fus%2Fproducts%2Fdiscovery-and-translational-research%2Fdna-rna-purification%2Fdna-purification%2Fgenomic-dna%2Fdneasy-blood-and-tissue-kit%2F&usg=AOvVaw2HPhb0J6RhMc_zq39I4VQq
- https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=&cad=rja&uact=8&ved=2ahUKEwiD_f2AjZP1AhUu7XMBHUpMAzQQFnoECAMQAQ&url=https%3A%2F%2Fwww.qiagen.com%2Fus%2Fresources%2Fdownload.aspx%3Fid%3D68f29296-5a9f-40fa-8b3d-1c148d0b3030%26lang%3Den&usg=AOvVaw3gG__v4c-QU-fPJWm34Fk1
- https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=&cad=rja&uact=8&ved=2ahUKEwj92vjNh5P1AhVqUGwGHYMdDQcQFnoECAMQAQ&url=https%3A%2F%2Fwww.colorado.edu%2Fecenter%2Fsites%2Fdefault%2Ffiles%2Fattached-files%2Fseracare_stability_of_genomic_dna_at_various_storage_conditions_isber2009.pdf&usg=AOvVaw1oa3mqlPVe0qSSfmh6HBo2
- https://easydna.com.au/knowledgebase/dna-testing-7-uses/
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