Navigation Links
UV Nicking of DNA in Agarose Gels for Enhanced Transfer and Detection of Megabase Size DNAs


Introduction
The transfer efficiency of large (>100 kb) DNAs from agarose gels to membrane is poor unless the DNAs are nicked prior to transfer. Although this has traditionally been accomplished with HCl depurination, UV irradiation (254 nm) provides a faster and more consistent alternative to HCl nicking. The GS Gene Linker UV chamber contains preprogrammed routines for many UV crosslinking and other controlled dosage applications, including UV nicking of pulsed field gels prior to transfer to membrane. The chambers unique sensor measures only the effective wavelengths of UV light, insuring reproducible results every time.


Results
High molecular weight DNA from a human HeLa cell line was cleaved with various restriction enzymes, resolved on the CHEF Mapper XA pulsed field electrophoresis system using the Auto Algorithm mode (50-750 kb), and irradiated with 60 mJoules of energy in the GS Gene Linker UV chamber after staining with ethidium bromide. The DNA was alkaline transferred to Zeta-Probe GT membrane, and probed with a human T cell receptor variable region (single copy gene) probe. As illustrated in Figure 1, bands ranging in size from 100 kb to greater than 700 kb were detected by the probe in the DNA lanes.


UV Nicking Protocol
Note: This protocol must be followed rigorously to obtain optimal results.
1. Following electrophoresis, stain the gel with 1.0 g/ml ethidium bromide (EtBr) for exactly 30 minutes with constant agitation. Use a fresh dilution of EtBr for each gel. Do not destain the gel.

2. Place the gel on a glass tray for tran
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
2. Recovery of DNA from LMP Agarose1
3. Advantages of ReadyAgarose Precast Gels Over 96-Well Handcast Gels for High-Throughput Analysis, Rev A
4. Enhanced PCR Cloning System
5. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
6. Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance
7. Enhanced siRNA Delivery and Long-term Gene Silencing
8. Measuring Intracellular Enhanced Green Fluorescent Protein With the VersaFluor Fluorometer
9. Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein
10. Increased Analyte Sensitivity through the Utility of Enhanced Mass-Resolution on the FinniganTSQ Quantum Discovery
11. Novel Bonding Chemistry Imparts Enhanced Polar Selectivity to TSK-GEL ODS-100V Reversed Phase Columns
Post Your Comments:
(Date:8/29/2014)... Inc. (NYSE MKT: PFNX), a clinical-stage biotechnology company ... to manufacture proteins including biosimilar therapeutics, today provided ... the second quarter ended June 30, 2014. ... public offering, we have the capital to continue ... our proprietary protein expression platform," stated Bertrand C. ...
(Date:8/29/2014)... are based on electrons, but one of the most ... i.e. circuits based on light (photons) instead of electrons. ... a stream of single photons and control their direction. ... attempts to achieve this control, but now scientists at ... steady stream of photons emitted one at a time ...
(Date:8/29/2014)... August 29, 2014 The global ... billion in 2019. It is expected to grow ... 2019, and was valued at $1.8 billion in ... Transparency Market Research. , For more information regarding ... . , The research report, titled “Companion Diagnostics ...
(Date:8/29/2014)... According to a new market report published ... market was valued at USD 3,754.6 million in 2012 and ... 2019at a CAGR of 5.9% from 2013 to 2019. , ... is afflicted by bone and joint disorders, and this number ... has and will continue to increase the demand for orthobiologics. ...
Breaking Biology Technology:Pfenex Reports Second Quarter 2014 Results and Provides Business Update 2Pfenex Reports Second Quarter 2014 Results and Provides Business Update 3Pfenex Reports Second Quarter 2014 Results and Provides Business Update 4Pfenex Reports Second Quarter 2014 Results and Provides Business Update 5Pfenex Reports Second Quarter 2014 Results and Provides Business Update 6Pfenex Reports Second Quarter 2014 Results and Provides Business Update 7Pfenex Reports Second Quarter 2014 Results and Provides Business Update 8Breakthrough in light sources for new quantum technology 2Companion Diagnostics Market to Hit $5.6 Billion in 2019: Transparency Market Research 2Companion Diagnostics Market to Hit $5.6 Billion in 2019: Transparency Market Research 3Global Orthobiologics Market to Reach $5,519.9 Million by 2019: Transparency Market Research 2Global Orthobiologics Market to Reach $5,519.9 Million by 2019: Transparency Market Research 3
...  Neurocrine Biosciences, Inc. (Nasdaq: NBIX ) announced today ... Neurocrine Biosciences, will be presenting at the 24th Annual Roth ... takes place on Tuesday, March 13 at 2:00 p.m. Pacific ... webcast and may be accessed on the Company,s website at ...
... ATLANTA, March 7, 2012 Harriet L. Robinson, Ph.D., ... a biotech company specializing in the development of HIV/AIDS ... scientists may be one step closer to a vaccine ... study results were unveiled by Dr. Robinson during a ...
... BETHESDA, Md., March 7, 2012   Spherix Incorporated ... innovator in biotechnology for therapy in diabetes, metabolic ... regulatory consulting services to food, supplement, biotechnology and ... its drug candidates, SPX-106T, arrested development and reduced ...
Cached Biology Technology:GeoVax Labs Presents New Findings on Potential HIV Vaccine at 2012 Conference on Retroviruses and Opportunistic Infections (CROI) 2GeoVax Labs Presents New Findings on Potential HIV Vaccine at 2012 Conference on Retroviruses and Opportunistic Infections (CROI) 3GeoVax Labs Presents New Findings on Potential HIV Vaccine at 2012 Conference on Retroviruses and Opportunistic Infections (CROI) 4SPX-106T Arrests Development of Atherosclerotic Plaques in Mouse Model of Cardiovascular Disease 2SPX-106T Arrests Development of Atherosclerotic Plaques in Mouse Model of Cardiovascular Disease 3SPX-106T Arrests Development of Atherosclerotic Plaques in Mouse Model of Cardiovascular Disease 4
(Date:8/29/2014)... made some surprising discoveries about the body,s initial ... (SIV), the team found that specialized cells in ... to viral invasion and are the source of ... beta (IL-1β). , Though aimed at the ... gut epithelium that provides a barrier to protect ...
(Date:8/29/2014)... Scientists at the Barshop Institute for Longevity and ... the UT Health Science Center at San Antonio, have ... the longest-lived rodent, the naked mole rat. , ... naked mole rats protects and alters the activity of ... proteins. , The factor also protects proteasome function in ...
(Date:8/29/2014)... recently, little has been known about what genetic changes ... of scientists, one of whom is a University of ... that genes controlling the development of the brain and ... The study was published Aug. 28 in Science ... is online at http://www.sciencemag.org/ ., The domestication of ...
Breaking Biology News(10 mins):The early cost of HIV 2Factor in naked mole rat's cells enhances protein integrity 2New research reveals how wild rabbits were genetically transformed into tame rabbits 2New research reveals how wild rabbits were genetically transformed into tame rabbits 3
... Researchers at the David Geffen School of Medicine at ... proteins known as sodium glucose co-transporters (SGLTs), which pump ... the treatment of chronic diarrhea via oral rehydration therapy, ... The solution of the SGLT structure will accelerate development ...
... have provided vital information to relief workers in Myanmar ... devastating Cyclone Nargis that hit the country on 2 ... the UN Office for the Coordination of Humanitarian Affairs ... Disasters, referred to as ,the Charter, for support by ...
... Want to slow the signs of aging and live longer? ... could be a promising strategy. Calorie restriction has ... and mice. While scientists do not know how calorie restriction ... that it slows aging by decreasing a thyroid hormone, triiodothyronine ...
Cached Biology News:UCLA researchers clarify function of glucose transport molecule 2Extended cyclone relief efforts aided from space 2Extended cyclone relief efforts aided from space 3Looking for the Founatain of Youth? Cut your calories, research suggests 2