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:8/3/2015)... 2015  MiMedx Group, Inc. (NASDAQ: MDXG ), ... and patent-protected processes to develop and market advanced products ... Sports Medicine, Ophthalmic and Dental sectors of healthcare, announced ... Annual Canaccord|Genuity Growth Conference in Boston, ... CEO, William C. Taylor , President and COO, ...
(Date:8/3/2015)... -- Merck KGaA, Darmstadt, Germany ,  a ... the healthcare, life science and performance materials sectors, announced ... and launch of promising new fullerene derivatives for the ... Germany , is pursuing many projects ... Novel derivatives: highly stable, easy to process and cost ...
(Date:8/3/2015)... 2015  Aethlon Medical, Inc. (Nasdaq: AEMD ), ... life-threatening diseases, today announced that it will host its ... on Thursday, August 13, 2015 at 4:30 p.m. Eastern ... the release of fiscal 2016 first quarter results.  A ... two hours after the call at the Investor Relations ...
(Date:8/3/2015)... Aug. 3, 2015   Intrexon Corporation (NYSE: ... half and second quarter 2015 financial results after the ... The Company will host a conference call at 5:30 ... general business update. The conference call may ... (International), and asking to join the "Intrexon Corporation Call."  ...
Breaking Biology Technology:MiMedx To Present At 35th Annual Canaccord|Genuity Growth Conference 2Merck KGaA, Darmstadt, Germany, and Nano-C Present Enhanced Materials for Organic Photovoltaics 2Merck KGaA, Darmstadt, Germany, and Nano-C Present Enhanced Materials for Organic Photovoltaics 3Merck KGaA, Darmstadt, Germany, and Nano-C Present Enhanced Materials for Organic Photovoltaics 4Aethlon Medical To Host First Quarter Fiscal 2016 Investor Call on August 13, 2015 2Intrexon to Announce First Half and Second Quarter 2015 Financial Results and Host Conference Call on August 10th 2
... Hollingsworth & Vose,Company (H&V), a leading ... introduced its HVision(TM) technology platform, a collaborative,process ... of,breakthrough products. "HVision will provide H&V ... a new generation of products that exceed ...
... Corporation (OTC,Bulletin Board: KDUS.OB) announced today financial results for the ... the third quarter of 2007 and for the same period,in ... $240,227, compared to,net income of $200,335 for the same period ... third quarter of 2007 and for the same period in,2006. ...
... Stock Exchange: EVT) Reported Today Results for,the First ... OXFORD, England, November 13 /PRNewswire-FirstCall/,-- Highlights: ... - Compelling results achieved in two proof-of-concept ... 201 - Sale of the Chemical Development ...
Cached Biology Technology:H&V Introduces HVision Technology Platform to Accelerate Development of Breakthrough Products 2Cadus Reports Third Quarter 2007 Results 2Cadus Reports Third Quarter 2007 Results 3Cadus Reports Third Quarter 2007 Results 4Evotec Reports Third Quarter 2007 Results 2Evotec Reports Third Quarter 2007 Results 3Evotec Reports Third Quarter 2007 Results 4Evotec Reports Third Quarter 2007 Results 5Evotec Reports Third Quarter 2007 Results 6Evotec Reports Third Quarter 2007 Results 7Evotec Reports Third Quarter 2007 Results 8
(Date:7/9/2015)... 9, 2015  Synaptics Inc. (NASDAQ: ... interface solutions, today announced a new Match-in-Sensor™ ... hardware encapsulated fingerprint sensor and matching solution. ... literally off the grid, isolating fingerprint image ... the fingerprint sensor to provide ultimate protection ...
(Date:7/7/2015)... 30, 2015 Research ... addition of the "Capacitive Fingerprint Sensors Patent ... To this date, fingerprint sensing technology is the ... sensors are well developed. This patent landscape focuses ... The domain of capacitive fingerprint sensors is ...
(Date:7/2/2015)... Fingerprint Cards has received an order for its ... distributor World Peace Industrial Group (WPI), part of WPG Holdings. ... . Deliveries are planned to take place during Q3 ... manufacturers in Asia . The order value ... of approximately 2,200 MSEK for 2015. Jörgen Lantto, ...
Breaking Biology News(10 mins):Synaptics Announces World's First Match-in-Sensor Fingerprint Authentication Technology 2Synaptics Announces World's First Match-in-Sensor Fingerprint Authentication Technology 3Worldwide Capacitive Fingerprint Sensors Patent Landscape Market Report 2015-2020 - Main IP Holders include Seiko Epson, AuthenTec, Sony and STMicroelectronics 2FPC Receives Order for Touch Fingerprint Sensors of 268 MSEK 2
... collective efforts to develop new drugs, the pharmaceuticals industry ... application of natural biological transformation processes. One of the ... cells in a bioreactor filled with a liquid culture ... regulations, the conditions inside the bioreactor need to be ...
... various Earth system science disciplines recently gathered to address ... in the future? Of course, possible answers to ... changing climatic conditions were to alter local vegetation, how ... Answering these questions with certainty would allow us to ...
... of BioScience, researchers present a sophisticated new ... ethanol on greenhouse gas emissions. The study, conducted by ... focuses on how mandated increases in production of the ... domestically and elsewhere. In response to the increased demand ...
Cached Biology News:Clean sampling from bioreactors 2Networking initiative to support interdisciplinary research 2Networking initiative to support interdisciplinary research 3More maize ethanol may boost greenhouse gas emissions 2
Rabbit polyclonal to Glutathione S Transferase alpha ( Abpromise for all tested applications). Antigen: Full length protein: Human Glutathione S Transferase alpha. Entrez GeneID: 2938 Swi...
Affinity Purified Rabbit Anti-mouse sRANKL Biotinylated Polyclonal Antibody...
Mouse anti-IkBa/MAD-3 Class: Antibody Product Group: Signalling molecules and phospho-specific Antibody...
Mouse monoclonal [002-49] to C1s ( Abpromise for all tested applications). entrezGeneID: 716 SwissProtID: P09871...
Biology Products: