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:1/14/2014)... As pet owners drew up their lists of New Year,s resolutions ... care of my furry companion." Nowadays there are pet salons, dog ... to take the little canine or feline darlings along wherever mommy ... some pricey toys at the pet store. But anyone who has ...
(Date:1/14/2014)... BETHESDA, Md. , Jan. 14, 2014 ... two institutes from the National Institutes of Health (NIH) ... safer, more effective treatments to patients on a faster ... Advancing Translational Sciences (NCATS) and the National Eye Institute ...
(Date:1/14/2014)... 2014  RXi Pharmaceuticals Corporation (OTCQX: RXII), a biotechnology ... therapies addressing major unmet medical needs using RNA-targeted ... of Allowance from the United States Patent and ... (sd-rxRNA®), for the treatment of fibrosis. The patent ...
(Date:1/14/2014)... 13, 2014 Bob Hainsey, a semiconductor ... 20 years in the industry, has joined SPIE, ... will serve as the society’s Science and Technology Strategist. ... SPIE as our Technology Strategist, further strengthening the technical ...
Breaking Biology Technology:New Year's Resolution: Give Pets the Gift of Top-Notch Health Care 2Organovo Announces Collaboration with National Institutes of Health 2Organovo Announces Collaboration with National Institutes of Health 3RXi Pharmaceuticals Receives US Notice of Allowance for a Key Patent Relating to its Self-Delivering Technology with sd-rxRNAs targeting CTGF, including RXI-109, for the Treatment of Fibrotic Disorders 2RXi Pharmaceuticals Receives US Notice of Allowance for a Key Patent Relating to its Self-Delivering Technology with sd-rxRNAs targeting CTGF, including RXI-109, for the Treatment of Fibrotic Disorders 3Photonics R&D Leader Bob Hainsey Joins SPIE Technical Staff 2
... has launched PATHT HTS, a new platform for high-throughput ... the microarray industry's annual conference, Chips To Hits in ... the performance of PATH protein microarray slides," said Robert ... BioSurfaces. "PATH HTS was designed to meet the ...
... The NeuronFarm, a developer of Web-based applications intended ... immediate availability of its flagship product, 3D-Readers. , ... third through eighth graders in several school studies, ... automated text scoring, engaging simulations and immediate feedback ...
... As more organizations look at what needs to be ... an area that has come under the spotlight is ... corporate governance issues have become issues even within the ... Act (SOX) has become a reason to organize and ...
Cached Biology Technology:NeuronFarm's 3D-Reader ready to ship 2Sarbox necessitating ECM software, but beware of `best practices' 2Sarbox necessitating ECM software, but beware of `best practices' 3
(Date:4/22/2014)... Consortium Ltd.,s nanotechnology expert will present a poster ... at the 7th International Nanotoxicology Congress to ... , Dr. Monita Sharma will outline a strategy consistent ... of Sciences, " Toxicity Testing in the 21st Century: ... of non-animal methods involving human cells and cell lines ...
(Date:4/22/2014)... presented Beaumont Health System with prestigious, competitive research ... athletic and nonathletic injuries. , The American ... grant to fund stem cell-based research that could ... ruptured knee anterior cruciate ligament, or ACL. According ... cruciate ligament injury is the over-stretching or tearing ...
(Date:4/22/2014)... at the University of Kentucky has discovered new methods ... , The research, led by Peixuan Guo, professor ... the UK College of Pharmacy and Markey Cancer Center, ... Boiling-Resistant Anionic Polymer Material To Build Robust Structures with ... and Daniel L. Jasinski. , The article, which will ...
Breaking Biology News(10 mins):Beaumont awarded grants to study sports-related injury treatment, improve recovery 2RNA shows potential as boiling-resistant anionic polymer material for nanoarchitectures 2
... has discovered the first identified droplets of spider blood in ... , Two droplets of blood, technically known as haemolymph, have ... - Filistatidae - a family commonly found in South America ... examples of spider blood ever found in an amber fossil. ...
... painkiller prompts brain to release endorphins, bringing real relief ... medicine will relieve pain is enough to prompt the ... painful sensations, a new University of Michigan study finds. ... the brain's own pain-fighting chemicals, called endorphins, play a ...
... completed 60 days of voluntary bedrest in order to ... body. The research team in Toulouse, France, is now ... continue and enhance the research. , At the beginning ... of the WISE study (Women International Simulation Experiment), were ...
Cached Biology News:Spider blood found in 20 million year old fossil 2Thinking the pain away? Study shows the brain's painkillers may cause 'placebo effect' 2Thinking the pain away? Study shows the brain's painkillers may cause 'placebo effect' 3Thinking the pain away? Study shows the brain's painkillers may cause 'placebo effect' 4Female volunteers prepare for a second 'bedrest' 2
Recombinant Rat Leptin, CF...
delta-like 1 homolog (Drosophila)...
... 71-18 mutS and ES1301 mutS are mismatch ... of these strains prevents repair of the ... high mutation efficiencies and making them helpful ... Altered Sites II Mutagenesis Systems. Both ES1301 ...
Request Info...
Biology Products: