Navigation Links
Genomic DNA Isolation From 1 ml of Body Fluid


AquaPure Genomic DNA Isolation Kit
Catalog #732-6340
Expected yield range: 250 g DNA (15-ml tube prep)


Method
Cell Lysis
1. Add 1 ml body fluid (e.g., cerebrospinal fluid, plasma, saliva, serum, sputum, synovial fluid, urine, whole blood, milk) to a sterile 15 ml centrifuge tube containing 5 ml cell lysis solution. Pipet up and down several times to mix thoroughly. Note: if the sample has a high protein content, 0.5 ml body fluid may be added to 5.5 ml cell lysis solution.

2. Heat to 65C for 15 min to complete lysis. Alternatively, for maximum yield, add 30 l of proteinase K (20 mg/ml) and incubate lysate at 55C for 1 hr to overnight.

RNase Treatment (Optional)
1. Add 30 l RNase A solution to the cell lysate.

2. Mix the sample by inverting the tube 25 times and incubate at 37C for 1560 min.

Protein Precipitation
1. Cool sample to room temperature.

2. Add 2 ml protein precipitation solution to the lysate.

3. Vortex sample at high speed for 20 sec to mix the protein precipitation solution uniformly with the lysate.

4. Place sample into an ice bath for 515 min.

5. Centrifuge at 2,000 x g for 10 min. The precipitated proteins should form a tight pellet. Note: if body fluid has a high lipid content, particulates may stay near top of tube; see alternative transfer method in step 1 below.

DNA Precipitation
1. Pour the supernatant containing the DNA (leaving behind the p recipitated protein pellet) into a clean 15 ml centrifuge tube containing 6 ml 100% isopropanol (2-propanol). Altern a t i v e l y, if p a rticulates are present, transfer supernatant using a pipet so that particulates are excluded. If the DNA yield is expected to be low (<20 g), add a DNA carrier such as glycogen (10 l of 20 mg/ml glycogen per 6 ml isopro p a n o l ) .

2. Mix the sample by inverting gently 50 times and keep tube at room temperature for at least 5 min.

3.Centrifuge at 2,000 x g for 10 min. The DNA may or may not be visible as a small white pellet, depending on yield.

4. Pour off the supernatant and drain tube briefly on clean absorbent paper. Add 6 ml 70% ethanol and invert the capped tube several times to wash the DNA pellet.

5. Centrifuge at 2,000 x g for 1 min. Carefully pour off the ethanol. Pellet may be loose, so pour slowly and watch pellet to ensure it stays in the tube.

6. I n v e rt and drain the tube on clean absorbent paper and allow to air- d ry for 15 min.

DNA Hydration
1. Add 100 l of DNA hydration solution (100 l will give a concentration of 200 g/ml if the yield is 20 g DNA).

2. Allow DNA to rehydrate overnight at room temperature . A l t e rn a t i v e l y, heat at 65C for 1 hr. Tap tube periodically to aid in dispersing the DNA.

3. Vo rtex briefly and pulse-spin before use. Store DNA at 28 C .


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
2. New Protocols for Isolating High- Molecular-Weight Genomic DNA
3. Easily Amplify Genomic DNA with Long-Distance PCR
4. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
5. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
6. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
7. Mouse Tail Genomic DNA Isolation Protocol(1)
8. Genomic DNA Isolation Protocol(1,2,3)
9. Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood Mini Kit
10. Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood Mini Kit
11. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/10/2017)... ... October 10, 2017 , ... Dr. Bob Harman, founder and CEO of ... Club. The event entitled “Stem Cells and Their Regenerative Powers,” was ... Dr. Harman, DVM, MPVM was joined by two human doctors: Peter B. Hanson, M.D., ...
(Date:10/10/2017)... Calif. , Oct. 10, 2017 SomaGenics ... from the NIH to develop RealSeq®-SC (Single Cell), expected ... for profiling small RNAs (including microRNAs) from single cells ... Program highlights the need to accelerate development of approaches ... "New techniques for measuring levels ...
(Date:10/10/2017)... (PRWEB) , ... October 10, 2017 , ... ... recipients of 13 prestigious awards honoring scientists who have made ... in a scheduled symposium during Pittcon 2018, the world’s leading conference and exposition ...
(Date:10/9/2017)... ... 09, 2017 , ... The award-winning American Farmer television series will feature 3 ... airs Tuesdays at 8:30aET on RFD-TV. , With global population estimates nearing ten ... to continue to feed a growing nation. At the same time, many of our ...
Breaking Biology Technology:
(Date:4/5/2017)... -- KEY FINDINGS The global market for ... of 25.76% during the forecast period of 2017-2025. The ... the growth of the stem cell market. ... INSIGHTS The global stem cell market is segmented on ... stem cell market of the product is segmented into ...
(Date:3/30/2017)... , March 30, 2017  On April 6-7, 2017, ... the Genome hackathon at Microsoft,s headquarters in ... competition will focus on developing health and wellness apps ... Hack the Genome is the first hackathon ... The world,s largest companies in the genomics, tech and ...
(Date:3/29/2017)... 2017  higi, the health IT company that operates ... America , today announced a Series B investment ... EveryMove. The new investment and acquisition accelerates higi,s strategy ... transform population health activities through the collection and workflow ... higi collects and secures data today on behalf of ...
Breaking Biology News(10 mins):