Navigation Links
Genomic DNA Isolation Protocol for 40 l Avian Whole Blood


AquaPure Genomic DNA Isolation Kit
Catalog #732-6340
Expected yield range: 100300 g DNA


Method
Cell Lysis
1. Use fresh blood within 10 sec of venipuncture to avoid clotting. Alternatively, collect whole blood in EDTA to prevent the blood from clotting and reduce DNA degradation. Blood may be either fresh or frozen. Store fresh blood sample at 28C for not more than 5 days to obtain optimal DNA isolation results.

2. Add 40 l whole blood to a 15 ml microfuge tube containing 6 ml cell lysis solution. Quickly pipet up and down 35 times to lyse the cells. Usually no incubation is required; however, if cell clumps are visible after mixing, incubate at 37C until the solution is homogeneous. Samples are stable in cell lysis solution for at least 18 months at room temperature.

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

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

Protein Precipitation
1. Cool sample to room temperature.

2. Add 2 ml protein precipitation polution to the RNase A treated cell lysate.

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

4. Centrifuge at 2,000 x g for 10 min. The precipitated proteins will form a tight pellet. If the protein pellet is not visible or is not tight, repeat step 3, incubate on ice for 5 min, then repeat step 4.

DNA Precipitation
1. Leaving behind the precipitated protein pellet, pour the supernatant containing the DNA into a 15 ml centrifuge tube containing 6 ml 100% isopropanol (2propanol).

2. Mix the sample by gently inverting the capped tube 50 times.

3. Centrifuge at 2,000 x g for 3 min; the DNA will be visible as a small white pellet.

4. Pour off the supernatant and drain tube 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 to ensure that pellet stays in the tube.

6. Invert and drain the tube on clean absorbent paper and allow to air-dry 15 min.

DNA Hydration
1. Add 500 l DNA hydration solution (500 l will give a concentration of 400 g/ml if the total yield is 200 g DNA).

2. Allow DNA to rehydrate overnight at room temperature. Alternatively, heat at 65C for 1 hr. Tap tube periodically to aid in dispersing the DNA.

3. Store DNA at 28C.


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:6/23/2016)... ... June 23, 2016 , ... Mosio, ... second eBook, “Clinical Trials Patient Recruitment and Retention Tips.” Partnering with experienced clinical ... eBook by providing practical tips, tools, and strategies for clinical researchers. , “The ...
(Date:6/23/2016)... 2016 Houston Methodist Willowbrook Hospital has ... Association to serve as their official health care ... Willowbrook will provide sponsorship support, athletic training services, ... coaches, volunteers, athletes and families. "We ... Association and to bring Houston Methodist quality services ...
(Date:6/23/2016)... ... June 23, 2016 , ... Supplyframe, the Industry ... Supplyframe Design Lab . Located in Pasadena, Calif., the Design Lab’s mission ... hardware projects are designed, built and brought to market. , The Design Lab ...
(Date:6/23/2016)... -- Andrew D Zelenetz , ... Published recently in Oncology & ... Andrew D Zelenetz , discusses the fact ... placing an increasing burden on healthcare systems worldwide, ... the patents on many biologics expiring, interest in ...
Breaking Biology Technology:
(Date:6/9/2016)... in attendance control systems is proud to announce the introduction of fingerprint attendance control ... right employees are actually signing in, and to even control the opening of doors. ... ... ... Photo - ...
(Date:6/3/2016)... Das DOTM (Department ... hat ein 44 Millionen $-Projekt ... einschließlich Personalisierung, Registrierung und IT-Infrastruktur, an Decatur ... Implementierung von Identitätsmanagementlösungen. Zahlreiche renommierte internationale Anbieter ... aber Decatur wurde als konformste und innovativste ...
(Date:6/2/2016)... June 2, 2016 Perimeter Surveillance ... Unmanned Systems, Physical Infrastructure, Support & Other Service  ... visiongain offers comprehensive analysis of the global ... will generate revenues of $17.98 billion in 2016. ... Inc, a leader in software and hardware technologies for ...
Breaking Biology News(10 mins):