Navigation Links
PFGE Analysis of Chromosomal DNA From a Single Hair Root

Contributed by Jacqueline Boultwood, Gail M. Abrahamson, and J. S. Wainscoat, Leukemia Research Fund, Molecular and Cytogenetic Haematology Unit, John Radcliffe Hospital, Headington, Oxford OX3 9DU, United Kingdom


Introduction
Tumor diagnoses, DNA fingerprinting, and other genetic analysis often require DNA obtained in a non-invasive manner. For example, peripheral blood may be used as a source from solid tumors, but not for haematological malignancies. We first reported the isolation and pulsed-field electrophoresis of genomic DNA obtained from a single hair root.1 Additional details are presented below.


Sample Preparation
One percent low melt agarose (SeaPlaque, FMC) was prepared in 1x PBS, 1 mM EDTA and added to a rectangular mold while molten. A single human hair was positioned in the middle of the block. After solidification, the block was placed in a NDS lysis solution consisting of 10 mM Tris, 0.5 mM EDTA, 1% sarcosyl, pH 9.5, 1 mg/ml Proteinase K, and 25 mM dithiothreitol. Blocks were incubated 24 hours at 50 C, with a fresh change of lysis buffer after 24 hours. Blocks were rinsed three times in NDS and stored at 4 C until required.

Enzyme digests were performed according to manufacturers directions. Prior to digestions, excess agarose was trimmed to within 2 mm of the hair.


Electrophoresis and Detection
The samples were resolved using the CHEF-DR II pulsed-field electrophoresis system. The gel was 1.0% agarose (SeaPlaque) in 0.5x TBE buffer chilled at 14 C. Electrophoresis was conducted at 200 volts for 24 hours with a 50-90 second switch time ramp.

Gels were first stained with ethidium bromide (Figure 1A
'"/>

Source:


Page: All 1 2

Related biology technology :

1. A Positive Selection Assay for Mutation Analysis in Big Blue Animals
2. Epitope-Tagging Vectors for Functional Analysis in Yeast
3. Map and Link Human Genetic Disorders with SSLP Analysis
4. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
5. Single-cell RT-PCR Analysis of Paramecium Primaurelia with the Eppendorf cMaster RTplusPCR System
6. Structural Analysis of Glycosylated Peptides in Complex Mixtures with Ion Trap MSn
7. Use of Transcriptor Reverse Transcriptase in Microarray Analysis
8. Increased Sensitivity in Microarray Analysis
9. Gene-Expression Analysis of TP, DPD, and TS Using the LightCycler mRNA Quantification KitsPLUS
10. Agilent 2100 Bioanalyzer Automated Analysis System
11. Analysis of Microarray Data
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: PFGE Analysis Chromosomal DNA From Single Hair Root

(Date:8/18/2014)... Aug. 18, 2014 BioElectronics Corporation ... consumer medical devices said that its EVP ... by CEOLIVE.TV as part of its Executive ... is available at http://ceolive.tv/bioelectronics-corp-otc-biel/biel-videos/2282-ceolive-tv-interview-bioelectronics-corp-otcqb-biel ... to investors the unique advantages of BioElectronics ...
(Date:8/18/2014)... UPTON, NYElectronic devices with unprecedented efficiency and ... materials that use built-in electric polarizations to ... magnets inside most popular data-driven technology. But ... stumbling blocks, including a curious habit of ... at the U.S. Department of Energy,s Brookhaven ...
(Date:8/18/2014)... YORK , August 18, 2014 ... published by Transparency Market Research "Life Science Reagents ... Flow Cytometry, Electrophoresis, Chromatography, In-Vitro Diagnostic Reagents and ... and Academic Institutions, Clinical Laboratories, and Forensic Laboratories)- ... Forecast, 2013 - 2019", the global life science ...
(Date:8/18/2014)... Aug. 18, 2014 TNI BioTech, Inc. (OTC-BB: ... the manufacturing and marketing of innovative therapies for autoimmune ... is providing a second notice of TNI BioTech,s annual ... the announcement, Noreen Griffin , founder and CEO ... for your participation in the Annual Meeting. On behalf ...
Breaking Biology Technology:BioElectronics EVP Dr. Deepak Kotak Interviewed on CEO Live 2BioElectronics EVP Dr. Deepak Kotak Interviewed on CEO Live 3Promising ferroelectric materials suffer from unexpected electric polarizations 2Promising ferroelectric materials suffer from unexpected electric polarizations 3Promising ferroelectric materials suffer from unexpected electric polarizations 4Global Life Science Reagents Market Expected to Reach USD 25.7 Billion in 2019: Transparency Market Research 2Global Life Science Reagents Market Expected to Reach USD 25.7 Billion in 2019: Transparency Market Research 3Global Life Science Reagents Market Expected to Reach USD 25.7 Billion in 2019: Transparency Market Research 4Global Life Science Reagents Market Expected to Reach USD 25.7 Billion in 2019: Transparency Market Research 5TNI BioTech Inc., provides 2nd Notice of Annual Meeting September 4, 2014 2TNI BioTech Inc., provides 2nd Notice of Annual Meeting September 4, 2014 3TNI BioTech Inc., provides 2nd Notice of Annual Meeting September 4, 2014 4
... in Wisconsin, educations effect on the state economy and ... were a few of the many topics covered yesterday ... , ,The panel first discussed the relation between the ... Wisconsin-Madison Chancellor John Wiley began by citing ...
... a lot of lively correspondence with Midwest technology job seekers, ... hanging onto their jobs by their fingernails. , ,They write ... their experiences with finding a job, getting promoted and managing ... write about really annoying things that their co-workers and supervisors ...
... child waits for Christmas. Today, PalmOne introduces the ... I buy?" conundrum. , ,I saw the new Treo six weeks ... (Yes, even I take a break from the schmoozing to sneak ... fell in love. This newest handheld communicator takes everything that ...
Cached Biology Technology:UW and business leaders outline need for educated workers 2UW and business leaders outline need for educated workers 3Midwest technology bosses should stop flaunting their wealth at work 2Midwest technology bosses should stop flaunting their wealth at work 3Midwest technology bosses should stop flaunting their wealth at work 4Christmas comes early with Treo 650 2Christmas comes early with Treo 650 3
(Date:8/20/2014)... seawater signals a relaxing trip to the shore. But ... comes with an environmental hitch. When certain sunblock ingredients ... become toxic to some of the ocean,s tiniest inhabitants, ... animals. Their study appears in the ACS journal ... David Snchez-Quiles point out that other than staying indoors, ...
(Date:8/20/2014)... rugs and plastic bags could one day be made ... petroleum, scientists are now reporting. The novel process they ... deal with its agricultural and plastic waste problems, appear ... Athanassiou, Ilker S. Bayer and colleagues at the Italian ... constantly growing. In 2012, its production reached 288 million ...
(Date:8/20/2014)... a colony of honeybees grows to about 4,000 members, ... cycle: the building of a special type of comb ... of experts from the Department of Neurobiology and Behaviour ... starts the reproductive cycle of honeybee colonies. The results ... Science of Nature . , Reproduction isn,t always ...
Breaking Biology News(10 mins):Worker bees 'know' when to invest in their reproductive future 2
... fruits and vegetables in ozone-enriched environments reduces spoilage explains ... Conference in Harrogate. Dr Ian Singleton explains how ozone ... as it leaves no residue on foods. It ... can be lost due to microbial spoilage. Dr Singleton ...
... The benefits of blueberry consumption have been demonstrated ... benefits derived from their high polyphenol content. Blueberries ... on everything from aging to metabolic syndrome. Recently, ... Denton, TX, examined whether blueberries could play a ...
... 2010 For years, scientists have thought of DNA as ... RNA transcription. Now, research led by scientists from the Florida ... also act to fine-tune the activity of certain proteins known ... it possible to design therapies that could activate specific genes ...
Cached Biology News:Ozone reduces fungal spoilage of fruits and vegetables 2Blueberries may inhibit development of fat cells 2Scripps Research scientists uncover new DNA role in modifying gene function 2Scripps Research scientists uncover new DNA role in modifying gene function 3
Endothelin-converting enzyme 1 (EC 3.4.24.71) (ECE-1). [Source:Uniprot/SWISSPROT;Acc:P42892] Antigen: Recombinant Protein Epitope Signature Tag (PrEST)....
...
Red Standard is sodium salt of resorufin....
X-Gal, 100 mg. 5-Bromo-4-chloro-3-indolyl-b-d-galactoside.Histochemical substrate for b-galactosidase, used in blue/white screening.Store at -20 C. Protect from moisture. Category: Nucleotides & Enzy...
Biology Products: