Navigation Links
Laser Capture Microdissection of muscle fiber populations and expression analysis by RT-PCR

cetylcholine receptor and slow MyHC are amplified. The housekeeping gene GAPDH was amplified as a reference for the amount loaded and the quality of the cDNA. All primers (Table II) are intron spanning to produce a cDNA specific PCR product. To avoid false positive results by PCR product carry-over, PCRs are setup in a flow hood.


17. 15 l of each PCR product were loaded on a 2% agarose gel and separated by 120 V for 25 minutes. Gels are stained with ethidium bromide and visualized using the Gel Doc 2000TM Gel Documentation System.


Results
The data presented in this application note demonstrates that it is possible to isolate a single muscle fiber type or a distinct muscle fiber population and analyze from it the expression of muscle specific genes by RTPCR. Figure 1 demonstrates the isolation of an individual fiber type by LCM. The ZenonTM technology based immunohistochemical staining of slow MyHC positive fibers gives a strong, unambiguous signal in less than 35 minutes. The acetylcholine esterase stain of Karnowsky and Roots allows us to assess rapidly the innervation pattern of muscle fibers. When used alone, this esterase stain can provide results within 5-10 minutes. Hence, rapid identification of distinct fibers based on their immunohistochemistry and innervation pattern allows us time to process tissue via LCM and isolate mRNA without significant time for degradation.

Using this type of analysis on limb muscle confirms data previously obtained by other means4. Hence, we can apply the technique to another muscle allotype, the EOM, and be sure that this protocol and the Arcturus PixCell II Laser Capture Microdissection System is sufficient to distinguish muscle fiber types and give new insight into the differences among muscle allotypes.


References
1. Fischer MD, Gorospe
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
2. Laser microdissection and nucleic acid purification - a Leica - QIAGEN cooperation
3. Working With Laser Capture Microdissection Samples
4. Laser Capture Microdissection (LCM) for the Analysis of Macrophage Gene Expression from Atherosclerotic Lesions
5. Laser Capture Microdissection (LCM) and Expression Profiling of Long-Range Projection Neurons
6. Differential Gene Expression Analysis of Pure Cell Populations from Frozen UterineTissue Using Laser Capture Microdissection (LCM) and cDNA Microarrays
7. Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein
8. Primary cells, rat heart muscle
9. High-throughput gene expression analysis to screen for anti-asthma drugs
10. Quantitative end-point RT-PCR gene expression measurement using the Agilent 2100 Bioanalyzer and standardized RT-PCR
11. Cytosolic expression of Green Fluorescent Protein (GFP) and its derivatives in the yeast Saccharomyces cerevisiae: Detection in vivo using the Varian Cary Eclipse
Post Your Comments:
(Date:8/28/2014)... A new research platform uses a laser ... structures undergoing stress and heating, an approach ... for microelectronics and batteries., This new technique, ... how heating and the surface stress of ... have discussed the merits of surface-stress influence ...
(Date:8/28/2014)... August 28, 2014 The ability for ... culture that will allow individuals to perform at the ... important for employers looking to compete in this post–recession ... embracing this trend, as the sector increasingly focuses on ... company, which will have real results on the organization's ...
(Date:8/28/2014)... 2014 Sterlitech is proud to announce ... its stable of products . These additional ... and include membrane filters with surface charges. ... demand for our membrane process testing equipment, and correspondingly, ... sheet membranes,” explains Sterlitech President Mark Spatz. “The ...
(Date:8/28/2014)... August 27, 2014 Scientists, researchers, and ... SPIE Laser Damage 2014 symposium. Marking its ... high-power lasers will run 14-17 September. The event is ... and photonics . , The premier conference for basic ... damage to optical materials will engage researchers and engineers ...
Breaking Biology Technology:New analytical technology reveals 'nanomechanical' surface traits 2New analytical technology reveals 'nanomechanical' surface traits 3Klein Hersh International Leads Philadelphia Area Life Science Industry in Company Culture 2Klein Hersh International Leads Philadelphia Area Life Science Industry in Company Culture 3Sterlitech Corporation Enhances Its Selection of Flat Sheet Membrane Filters 2Laser Damage to Draw Researchers, Engineers, Scientists to 46th Annual Conference 2Laser Damage to Draw Researchers, Engineers, Scientists to 46th Annual Conference 3
... Inc. and flexcipio s.a. are pleased to ... Clinical Conductor Clinical Trial Management System (CTMS) and ... aligns the industry,s most comprehensive enterprise CTMS, with ... the best tools to optimize efficiency, productivity, and ...
... April 14 /PRNewswire/ - ImmunoVaccine Technologies Inc. (IVT), ... of a 3-year research agreement with Defence Research ... the preclinical application of IVT,s DepoVax(TM) vaccine formulation ... prevent, prepare for, and respond to biological threats. ...
... Showing Statistically Significant Improvement in Overall Survival -- First ... Data to be Presented at Plenary Session at Upcoming ... Call Today at 9:00 AM ET -SEATTLE, April 14 ... today that the pivotal Phase 3 IMPACT study of ...
Cached Biology Technology:Bio-Optronics Adds Innovative Electronic Data Capture (EDC) Solution to Clinical Conductor Clinical Trial Management System (CTMS), Continues to Improve Workflow and Productivity 2ImmunoVaccine Technologies Collaborates with Defence Research & Development Canada to Strengthen Canada's Ability to Deal with Biological Threats 2PROVENGE Significantly Prolongs Survival in Men With Advanced Prostate Cancer in Pivotal Phase 3 IMPACT Study 2PROVENGE Significantly Prolongs Survival in Men With Advanced Prostate Cancer in Pivotal Phase 3 IMPACT Study 3PROVENGE Significantly Prolongs Survival in Men With Advanced Prostate Cancer in Pivotal Phase 3 IMPACT Study 4
(Date:8/28/2014)... traditional cigarettes, but still release toxins into the ... , Scientists studying secondhand smoke from e-cigarettes discovered ... particles, with close-to-zero exposure to organic carcinogens. However, ... second-hand e-cigarette smoke were found to be significantly ... of polycyclic aromatic hydrocarbons cancer-causing organic compounds ...
(Date:8/28/2014)... and his collaborators have developed an online analytic tool ... re-engineering cells for biomedical investigation. CellNet is a free-use ... stem cell engineering. Details of CellNet and its application ... papers in the journal Cell . , "This ... all types of cell-based investigations and can potentially offer ...
(Date:8/28/2014)... MD FASEB MARC (Maximizing Access to Research ... for the Genetics Society of America,s 27th Annual ... October 3, 2014 in Pacific Grove, California. ... of students, post doctorates and scientists from underrepresented ... community and to encourage the participation of young ...
Breaking Biology News(10 mins):Second-hand e-cig smoke compared to regular cigarette smoke 2New tool aids stem cell engineering for medical research 2
... ITHACA, N.Y. When it comes to potentially doubling the ... may be in the spit. Researchers at Cornell University, ... of Colombia, have discovered that when a major South American ... The secret to this increased yield, they write in the ...
... Mich. Solar power might be nature,s most plentiful and ... lure water insects away from critical breeding areas, a Michigan ... or other methods to break up the polarized reflection of ... likely to deposit eggs on the panels thinking that they ...
... and others interested in the chemical sciences are invited to ... (ACS) Webinars, focusing on tips on how to improve your ... Thursday, June 3, from 2 3 p.m. EDT, the ... for "Effective Technical Writing Short Course" offered by the ACS ...
Cached Biology News:To double spud production, just add a little spit 2Solar panels can attract breeding water insects 2
Recombinant Porcine IL-1 alpha/IL-1F1, CF...
... Immunogen KLH-conjugated, synthetic ... 54-75 (RRGPHRNPG-PGDDDGQRSRSSS) of mouse p19ARF. ... to amino acids 62-75. Clone ... Routinely evaluated by immunoblot. ...
... siSolutions siRNA Custom Synthesis ... effective duplex RNA to knockdown ... 21-mers (19 RNA base pairs ... recently-published 27-mers, or other lengths ...
Request Info...
Biology Products: