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:6/24/2016)... TOKYO , June 24, 2016  Regular discussions on ... to take place between the two entities said Poloz. ... in Ottawa , he pointed to the ... and the federal government. ... Poloz said, "Both institutions have common economic goals, why not ...
(Date:6/23/2016)... , ... June 23, 2016 , ... ... release of its second eBook, “Clinical Trials Patient Recruitment and Retention Tips.” Partnering ... retention in this eBook by providing practical tips, tools, and strategies for clinical ...
(Date:6/23/2016)... , June 23, 2016 A person commits ... the crime scene to track the criminal down. ... U.S. Food and Drug Administration (FDA) uses DNA evidence to ... Sound far-fetched? It,s not. The FDA has ... to support investigations of foodborne illnesses. Put as simply as ...
(Date:6/23/2016)... , June, 23, 2016  The Biodesign Challenge ... envision new ways to harness living systems and biotechnology, ... Art (MoMA) in New York City ... 130 participating students, showcased projects at MoMA,s Celeste Bartos ... Paola Antonelli , MoMA,s senior curator of architecture and ...
Breaking Biology Technology:
(Date:6/2/2016)... Perimeter Surveillance & Detection Systems, ... Infrastructure, Support & Other Service  The latest ... comprehensive analysis of the global Border Security market ... of $17.98 billion in 2016. Now: In ... in software and hardware technologies for advanced video surveillance. ...
(Date:5/24/2016)... facilitates superior patient care by providing unparalleled technology to leaders of the medical imaging ... product recently added to the range of products distributed by Ampronix. Photo ... ... ... News ...
(Date:5/16/2016)... 2016   EyeLock LLC , a market leader ... of an IoT Center of Excellence in ... development of embedded iris biometric applications. EyeLock,s ... and security with unmatched biometric accuracy, making it the ... DNA. EyeLock,s platform uses video technology to deliver a ...
Breaking Biology News(10 mins):