Navigation Links
Detection of p53 Gene in Breast Cancer by Denaturing Gradient Gel Electrophoresis and the DCode System

ples obtained from breast cancers were examined from exon 5, 6, 7 and 8 of the p53 gene. They were amplified by PCR with a pair of primers which contains a 40 bp GC clamp in one of them.2 All products were analyzed both in perpendicular gradient gels and parallel gradient gels.

PERPENDICULAR GRADIENT GEL ANALYSIS
A 7.5 x 10 cm, 1 mm thick, 25% to 65% denatured gradient gel was made using 6% aclylamide/bis (37.5:1) in 1x TAE buffer (50 mM Tris, 25 mM acetic acid, 1.25 mM EDTA). Four different PCR products each from exon 5, 6, 7 and 8 were mixed in a 25 l volume and the total sample volume made up to 100 l. We added 100 ml 2x gel loading dye (70% glycerol, 0.05% bromophenol blue, 0.05% xylene cyanol, 2 mM EDTA) to these samples, and electrophoresed them on the DCode system at 130 V for 2.5 hours at 56 C. After electrophoresis, the gels were stained in a 1:10,000 dilution of SYBR Green I (Molecular Probes) in 1x TAE buffer for 45 minutes and destained in 1x TAE buffer for 45 minutes. The gels were imaged under UV transillumination.

PARALLEL GRADIENT GEL ANALYSIS
A 16 x 16 cm, 1 mm thick, 45% to 60% denatured gradient gel was made using 6% aclylamide/bis (37.5:1) in 1xTAE buffer. Five ml of PCR products (approximately 200300 ng) was mixed with 5 l 2x gel loading dye and electrophoresed on the DCode system at 150 V for 3.5 hours at 60 C. The post-run analysis was the same as perpendicular gradient gel analysis.


Results and Discussion

The optimum concentration of denaturant to be used for parallel gradient gels was determined from the perpendicular gradient gel analysis. All exons separated and had a melting transiti
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A New PCR-based Mycoplasma Detection Method
3. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
4. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
5. Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes
6. Gene Expression Arrays: Highly Sensitive Detection of Expression Patterns with Improved Tools for Target Amplification
7. The DIG System Nonradioactive and Highly Sensitive Detection of Nucleic Acids
8. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
9. Quantification of Nucleosomes in Serum by the Cell Death Detection ELISAplus
10. A Further Step in Understanding Apoptosis Direct Detection of PARP Cleavage
11. In Situ Cell Death Detection Kit
Post Your Comments:
(Date:1/15/2014)... Jan. 15, 2014 ­ RedBrick Health , a ... technology, today announces that EmblemHealth , the ... company, is now providing the RedBrick Compass health assessment, ... to all of its members. EmblemHealth is among the ...
(Date:1/15/2014)... January 15, 2014 A study has been ... the Formula 1 track could help to tackle the problem ... (MAT), Stowhealth (a GP surgery based in Stowmarket) and academics ... Simplyhealth. Telemetry technology, which is inspired by equipment ...
(Date:1/15/2014)... VA (PRWEB) January 15, 2014 Two ... have continued their sponsorship of an annual competition for ... in, and access to, innovative STEM study. The competition ... TEAMS: Tests of Engineering Aptitude, Mathematics, and Sciences ...
(Date:1/15/2014)... 15, 2014 DTS Language Services, Inc ... Web Portal for Life Science organizations who need document translations. ... subject matter of their documents in advance with a selection ... time-to-delivery of translations, often a critical factor in clinical and ...
Breaking Biology Technology:EmblemHealth Selects RedBrick Health to Power its Next-Generation Health Engagement and Behavior Change System 2EmblemHealth Selects RedBrick Health to Power its Next-Generation Health Engagement and Behavior Change System 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 2Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 4Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 2Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 3Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 4Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 5DTS Improves Efficiency for Life Science Document Translations 2
... announced today that it has become an approved provider ... Care Centers of America (LCCA). ReBound™ is ... of life by accelerating the healing process and relieving ... Ease of use, portability, safety and the ability ...
... Growth Hormone Data Selected for Presentation at 5th International... -- NES-ZIONA, Israel, Sept. 27 ... ... --> ... var s = ,; var first_result; // Results are keyed by longUrl, so we need to grab ...
... 27 Clinical Genomics Pty. Ltd (Sydney, Australia) ... Australia) today announced that they have entered into ... discovered by scientists at the Garvan Institute. Under ... worldwide exclusive rights to the Garvan,s proprietary cancer ...
Cached Biology Technology:ReGear Life Sciences Partners with Life Care Centers of America 2PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 2PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 3PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 4PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 5PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 6PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 7PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 8PROLOR Biotech's Longer-Acting Human Growth Hormone Data Selected for Presentation at 5th International Congress Of The Growth Hormone Research Society and IGF Society 9Clinical Genomics Pty Ltd and The Garvan Institute of Medical Research Enter into Licensing Agreement 2
(Date:4/17/2014)... 16, 2014. Kessler Foundation has been named awardee ... the Department of Defense Spinal Cord Injury Research ... for the randomized, double-blinded, controlled, multi-site clinical trial, ... muscle strength after spinal cord injury. Dr. Forrest ... Research at Kessler Foundation. Two additional sites will ...
(Date:4/17/2014)... one per eight hundred births, Down syndrome - or ... of intellectual disability. It results from a chromosomal abnormality ... of chromosome 21 (1% of the human genome). A ... the Department of Genetic Medicine and Development at the ... Nature , shed light on how the extra ...
(Date:4/17/2014)... day about eight years ago, Katia Silvera , a ... father were on a field trip in a mountainous area ... had never seen before. , Unable to identify it, they ... orchid turned out to be an unnamed species. So Carnevali ... , "Lophiaris" is the genus name, comprising about 40 species ...
Breaking Biology News(10 mins):Kessler Foundation awarded Department of Defense grant for spinal cord injury research 2Trisomy 21: How an extra little chromosome throws the entire genome off balance 2Trisomy 21: How an extra little chromosome throws the entire genome off balance 3Orchid named after UC Riverside researcher 2Orchid named after UC Riverside researcher 3
... Thursday, July 12, 2012, Cleveland: The National Heart, Lung, ... grant to researchers at Cleveland Clinic,s Lerner Research Institute ... on the small-molecule byproducts of metabolism for ... cardiovascular diseases. "Despite the identification of numerous ...
... harsh conditions; they range over large areas of cold ... suggests wolverines take advantage of the crevices and boulders ... cover to cache and "refrigerate" food sources such as ... and other food collected during more plentiful times of ...
... UC Santa Barbara scientists trying to control the invasive ... in the form of a rapidly evolving beetle that ... consume the noxious weed. Their findings, in a paper ... enables range expansion of Diorhabda carinulata , a ...
Cached Biology News:Cleveland Clinic researchers receive $5 million grant to discover novel pathways to heart disease 2Study: Wolverines need refrigerators 2Tamarisk biocontrol efforts get evolutionary boost 2Tamarisk biocontrol efforts get evolutionary boost 3
... Engineered with new technology incorporating the ... unsurpassed flexibility, the new Varian NMR ... spectrometer available today., ,The Varian NMR ... transmitter and receiver channel, providing pulse ...
... cell labeling mix, 92.5 MBq, ... stabilized with 0.1% 2-mercaptoethanol and ... acid. *Amino acid mixture for ... l-[35S]Methionine and l-[35S]Cysteine. * > ...
... MMP-3, Human, Assay, 96 wells. ELISA.Specific ... and complexed MMP-3. No cross-reactivity with MMP-1, ... ng/ml.Suitable for use with cell culture supernatant, ... at -15 to -30 C. Category: ...
... for serial preparation of ... including cytosol/particulate/cytoskeleton/nuclear fractions from ... fractions can directly be ... such as 2-D gel/Western ...
Biology Products: