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:8/28/2015)... Aug. 28, 2015 /PRNewswire/ - RepliCel Life Sciences ... medicine company focused on the development of autologous ... quarter ended June 30, 2015, corporate highlights, and ... report are available at www.sedar.com ,  ... "RepliCel expects to launch its RCS-01 ...
(Date:8/27/2015)... ... ... Capital Edge Consulting , a nationwide consulting firm that specializes in helping ... one of the top 5000 companies listed on the 2015 Inc. 5000 , ... is truly an honor to be recognized as one of America’s fastest growing companies. ...
(Date:8/27/2015)... Calif. , Aug. 27, 2015  Asterias Biotherapeutics, ... the emerging field of regenerative medicine, today announced that ... conferences in the month of September.  ... present at Rodman & Renshaw,s 17th Annual Global Investment ... PT at the St. Regis Hotel in ...
(Date:8/27/2015)... ... , ... Inc. Magazine released its annual list of the 5,000 fastest-growing privately ... for the three years through 2014. Being named to the Inc. 5000 list designates ... country. , “We are thrilled to make the Inc. 5000 list for 2015,” ...
Breaking Biology Technology:RepliCel Reports Second Quarter 2015 Financial Results 2RepliCel Reports Second Quarter 2015 Financial Results 3RepliCel Reports Second Quarter 2015 Financial Results 4RepliCel Reports Second Quarter 2015 Financial Results 5Capital Edge Consulting Makes Inc. 5000 List of America’s Fastest-Growing Private Companies 2Capital Edge Consulting Makes Inc. 5000 List of America’s Fastest-Growing Private Companies 3Asterias Biotherapeutics to Present at Upcoming Investor Conferences 2iLab Solutions Named to 2015 Inc. 5000 List 2iLab Solutions Named to 2015 Inc. 5000 List 3
... N.J., Feb. 6 Senesco,Technologies, Inc. ("Senesco" or ... results of preclinical animal studies focused on multiple ... the University of Waterloo.,The goal of these studies, ... test the ability of Senesco,s Factor 5A technology ...
... a unique move to provide the,unparalleled fertility and ... Bay demand and deserve, The American Fertility,Association takes ... this Sunday,Feb. 10th for the first-ever West Coast ... fertility education and advocacy nonprofit,that embeds inclusiveness in ...
... FRANCISCO, Calif., Feb. 6 Rigel,Pharmaceuticals, Inc. (Nasdaq: ... chairman and chief executive officer, is scheduled to present,a ... 2008 in New York on,Monday, February 11 at 11:45 ... or the subsequent archived recording,log on to http://www.rigel.com ...
Cached Biology Technology:Senesco Technologies Reports on Results of Preclinical Multiple Myeloma Studies 2Senesco Technologies Reports on Results of Preclinical Multiple Myeloma Studies 3The American Fertility Association Brings Fertility Education to San Francisco or Parent Hopefuls - Straight and Gay 2
(Date:8/18/2015)... , Aug. 18, 2015  Navitas ... key contributor to the Submission Content Management ... IRISS, a non-profit forum. Dedicated to the Implementation ... a neutral platform for industry, vendors, health ... information and work towards a standard method ...
(Date:8/12/2015)... MINNETONKA, Minn. , Aug. 12, 2015   ... that supports the entire spectrum of clinical research, is ... ™ , the company,s comprehensive SaaS-based eClinical technology platform, ... growth during the first two quarters of 2015.   ... and second largest quarters measured by contract value sold ...
(Date:8/12/2015)... , Aug. 12, 2015  Synaptics Inc. ... human interface solutions, today announced that four of ... Match-in-Sensor solution, have officially been named FIDO Certified™ ... part of the certification, Synaptics, Natural ID™ fingerprint ... Universal Authentication Framework (UAF) standard and are interoperable ...
Breaking Biology News(10 mins):Navitas, a Key Contributor in IRISS Forum Webinar 2Navitas, a Key Contributor in IRISS Forum Webinar 3MedNet Solutions Experiences Explosive Corporate Growth 2Synaptics Announces FIDO Certification of Natural ID Fingerprint Solutions 2Synaptics Announces FIDO Certification of Natural ID Fingerprint Solutions 3
... Gil and Rongchao Jin have successfully used NMR ... which could advance the development and use of ... Their approach offers a significant advantage over routine ... determine whether the nanoparticles exist in a both ...
... made up of neurons make decisions using strikingly similar mechanisms, ... Science . In previous work, Cornell University biologist ... perform "waggle dances" to prompt other scout bees to inspect ... the new study, Seeley, a professor of neurobiology and behavior, ...
... Karanth of Bangalore, India, is the recipient of the ... first wild tigers at the age of 2 at ... Ullas Karanth. Krithi Karanth works on human dimensions of ... people-park relationships., With the National Geographic grant, she will ...
Cached Biology News:Carnegie Mellon researchers use NMR to determine whether gold nanoparticles exhibit 'handedness' 2Carnegie Mellon researchers use NMR to determine whether gold nanoparticles exhibit 'handedness' 3Young conservation biologist receives National Geographic's 10,000th grant 2
2,4,6, Trinitrophenyl hapten is conjugated to lipopolysaccharide (LPS)....
SHEEP ANTI BOVINE LACTOPEROXIDASE...
... and flexibility press-to-seal silicone isolators can be ... researcher. They are available either pre-cut to ... that can be easily trimmed to prepare ... non-cytotoxic and can be used to isolate ...
Request Info...
Biology Products: