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:9/17/2014)... 2014 Hussey Copper , a leader ... antimicrobial copper, announces Gilmour Academy Ice Arena,s conversion of ... antimicrobial solid touch surface to help protect against bacteria.  ... surface that kills greater than 99.9% of MRSA, E-coli ... , Innovative ...
(Date:9/17/2014)... FRANCISCO, Calif. , Sept. 17, 2014 ... a molecular diagnostics company pioneering the field of molecular ... to chief operating officer. Mr. Hall joined ... the company,s commercial entry into endocrinology. Since 2012, he ... as well as managed care and billing functions. As ...
(Date:9/17/2014)... Beckman Coulter , a global leader ... renewed its agreement with Personify for an ... Personify will continue to streamline the Beckman Coulter talent ... including time-to-fill, time-to-source and a deep reduction to cost-per-hire. ... was based on the success of our partnership over ...
(Date:9/17/2014)... NASA’s MESSENGER has just celebrated its 10 ... to have one of its latch valves, V27200-818, operating ... scientific investigation of the planet Mercury. It is only ... has provided a vast amount of information and images ... valves utilize solenoids that also include permanent magnets to ...
Breaking Biology Technology:Innovative Gilmour Academy Ice Arena changes more than 200 fixtures to Hussey Copper's MD-Cu29 antimicrobial copper to help protect against bacteria 2Innovative Gilmour Academy Ice Arena changes more than 200 fixtures to Hussey Copper's MD-Cu29 antimicrobial copper to help protect against bacteria 3Veracyte, Inc. Announces Promotion of Christopher M. Hall To Chief Operating Officer 2Veracyte, Inc. Announces Promotion of Christopher M. Hall To Chief Operating Officer 3Veracyte, Inc. Announces Promotion of Christopher M. Hall To Chief Operating Officer 4Beckman Coulter Signs 3-year RPO Contract with Personify 2Beckman Coulter Signs 3-year RPO Contract with Personify 3
... China, Dec. 15 /PRNewswire-Asia/ -- China Medicine ... "the Company"), a leading,developer and distributor of ... medicines (TCM), nutritional and dietary supplements,medical devices ... from,the Guangdong Food & Drug Administration to ...
... to Support Phase 3 Combination Rule Trial for NDA ... Australia and BEDMINSTER, N.J., Dec. 15 QRxPharma Limited ... company focused on the development and commercialization of therapies ... today initiation of a comparative study to evaluate the ...
... Platform Brings System Flexibility and Efficiency to the Advanced ... Scanning Systems , , ... confocal microscope and software package available, Nikon Instruments, Inc.,s ... scanning systems with dedicated NIS-Elements C software brings biological ...
Cached Biology Technology:China Medicine to Distribute Dental and Surgical Instruments 2China Medicine to Distribute Dental and Surgical Instruments 3QRxPharma Initiates Comparative Study for MoxDuoIR(TM) Dual-Opioid(TM) Pain Therapy 2QRxPharma Initiates Comparative Study for MoxDuoIR(TM) Dual-Opioid(TM) Pain Therapy 3Nikon Instruments A1 Confocal Laser Microscope Series With NIS-Elements C Software Delivers Fully Integrated Comprehensive Confocal Imaging Capabilities 2Nikon Instruments A1 Confocal Laser Microscope Series With NIS-Elements C Software Delivers Fully Integrated Comprehensive Confocal Imaging Capabilities 3
(Date:9/16/2014)... Health at the Mailman School of Public Health are ... and prenatal exposure to two phthalates used in a ... the journal Environmental Health Perspectives . , ... levels of the chemicals, butylbenzyl phthalate (BBzP) and di-n-butyl ... increase in risk of developing asthma between age 5 ...
(Date:9/16/2014)... Ohio (September 17, 2014)The North American Menopause Society ... the comprehensive care of midlife womenon everything from ... "The North American Menopause Society Recommendations for Clinical ... in the Society,s journal Menopause . This ... for the care of midlife women freely available ...
(Date:9/16/2014)... the "molecular brakes" that time the generation of important ... "hair cells" translate sound waves into electrical signals that ... sounds. If the arrangement of the cells is disordered, ... research will be published in The Journal of ... and Hey2 act as brakes to prevent hair cell ...
Breaking Biology News(10 mins):Phthalates heighten risk for childhood asthma 2Phthalates heighten risk for childhood asthma 3Hey1 and Hey2 ensure inner ear 'hair cells' are made at the right time, in the right place 2
... Virginia Commonwealth University immunologists studying mast cells, known to ... have identified a hormone-like molecule that can kill these ... mice. , The findings move researchers another step ... and may help researchers develop new treatments for allergy ...
... day, tens of thousands of biologists around the globe ... online algorithm called BLAST ?typically submitting 200 to 400 ... matched against the billions of letters for known genes. ... or evolutionary relationships, scientists routinely use BLAST to churn ...
... captures pictures beneath the living brain's surfaceResearchers at Stanford ... that can capture micron-scale images from deep in the ... combines a pair of powerful optical and mechanical techniques ... the hand. The results appear in the September 1, ...
Cached Biology News:VCU study shows hormone-like molecule kills cells that cause inflammation in allergic disease 2Drawing with DNA: 'Bioart' illuminates genomics 2Drawing with DNA: 'Bioart' illuminates genomics 3Microscopic brain imaging in the palm of your hand 2Microscopic brain imaging in the palm of your hand 3
Our original highly-purified, robust Taq Polymerase which delivers consistently high yields and minimal background...
A formulation of our Biolase DNA Polymerase which contains a Red Loading Dye...
... immunosorbent assay (ELISA) test kits are ... for the detection of drugs and/or their ... intended for forensic use only. It is ... by a quantitative method such as GC/MS. ...
... SUPERaseIn RNase Inhibitor (patent pending) is ... that noncovalently binds and inhibits the most ... B, C, 1 and T1. SUPERaseIn can ... contamination could be problematic. It is ideal ...
Biology Products: