Navigation Links
Laser Capture Microdissection of Cells Labeled with Enhanced Green Fluorescent Protein

d or fluorescence) during LCM allows accurate capture of EGFP-expressing cells

7. After completing LCM, perform extraction, purification and amplification of RNA using the Arcturus PicoPure and RiboAmp kits according to the manufacturers protocols.

8. Synthesize cDNA from amplified RNA with random hexamer primers and Superscript II reverse transcriptase to confirm expression of cell type-specific gene targets by quantitative real-time PCR (Bhattacherjee et al., 2004).


Results
During dissection, embryos in a litter that express EGFP in a pattern specific to neural crest cells can be identified by fluorescent stereo microscopy prior to fixation and embedding in OCT (Figure 1). EGFP-labeled neural crest cells can be identified and captured by LCM from partially fixed cryosections of embryonic tissue (Figure 2). Quantitative Real-Time PCR of the RNA extracted from these captured cells resulted in amplification of neural crest cell gene markers (Crabp1, Dlx5, Msx1, Slug), while expression of Engrailed-1 (En1), a gene that is not expressed in the neural crest cells was not detected (Table 1). These results demonstrate the utility of LCM to isolate specific EGFP-expressing cell populations from tissue sections of transgenic mouse embryos that can be used for subsequent molecular analyses.



1Expression of neural crest gene markers was determined using TaqMan quantitative realtime PCR.
2+ indicates detectable levels of gene expression; - indicates undetectable levels.
3Neural crest cDNA samples were prepared and subjected to QRT-PCR for each target gene in triplicate; mean Ct values are reported.
4Negative methodological control reactions, which lacked reverse transcriptase, did not amplify any detectable product.
5
'"/>

Source:


Page: All 1 2 3 4 5

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 muscle fiber populations and expression analysis by RT-PCR
8. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
9. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
10. Cell Proliferation Kit Outperforms the Competition in the Range of Cells Typically Counted
11. XL10-Gold Cells Supplied in New Single-Reaction Format
Post Your Comments:
(Date:9/18/2014)... -- MEI Pharma, Inc. (Nasdaq: MEIP ), an oncology ... cancer, announced today that Daniel P. Gold , Ph.D., ... NewsMakers in the Biotech Industry conference on Friday, September ... Broadway Hotel & Conference Center in New York ... accessed at www.meipharma.com . A replay will be available ...
(Date:9/18/2014)... 18 septembre 2014 Mapi, société créée ... 1974, a été depuis 40 ans l,un ... cliniques traditionnelles par des évaluations recentrées sur ... les impacts économiques des traitements. ... A cours d,une récente conférence téléphonique ...
(Date:9/18/2014)... SeqLL Inc., the owner of ... today the closing of a $1M Series-A funding ... Diagnostic Technologies, will increase the power of tSMS™ ... in March 2013 by Daniel Jones, a former ... BioSciences, SeqLL has continued to offer Helicos’ sequencing ...
(Date:9/18/2014)... sweeteners, promoted as aids to weight loss and diabetes ... and metabolic disease; and they do it in a ... the gut microbiota the substantial population of bacteria ... experiments in mice and humans, were published today in ... Elinav of the Weizmann Institute,s Immunology Department, who led ...
Breaking Biology Technology:MEI Pharma To Present At BioCentury's NewsMakers Conference 2MEI Pharma To Present At BioCentury's NewsMakers Conference 31974 - 2014 - Mapi fête ses 40 ans 2SeqLL Closes Series-A Funding Round for Expansion of Single Molecule Sequencing 2Gut bacteria, artificial sweeteners and glucose intolerance 2Gut bacteria, artificial sweeteners and glucose intolerance 3
... Sweden, July 14 PharmaSurgics AB.,successfully completes Phase ... is safe and well tolerated. Initially developed to ... combines,the pharmacological effect of PXL01 with the physical ... to be the first anti-adhesion drug on the,market. ...
... LA JOLLA, Calif., July 14 Synthetic Genomics ... solutions to address a variety of global challenges including ... and development agreement with ExxonMobil Research and Engineering Company ... algae. , , ...
... , ... today announced clearance for marketing by the United States Food and Drug Administration (K083292) ... , ... 14, 2009 -- EMcision Ltd. today announced clearance for marketing by the United States ...
Cached Biology Technology:Novel Product for the Prevention of Post-Surgical Adhesion Formation Completes Phase I Clinical Study 2Synthetic Genomics Inc. and ExxonMobil Research and Engineering Company Sign Exclusive, Multi-Year Agreement to Develop Next Generation Biofuels Using Photosynthetic Algae 2Synthetic Genomics Inc. and ExxonMobil Research and Engineering Company Sign Exclusive, Multi-Year Agreement to Develop Next Generation Biofuels Using Photosynthetic Algae 3EMcision Announces 510(k) Clearance of the Habib EndoHPB, its Novel Bipolar RF Device for the Palliation of Biliary and Pancreatic Tumours 2
(Date:9/17/2014)... four-year project, delivering its first release in 2004. After ... several EC projects, as a part of the EC-FP7 ... to increase the general awareness of the work done ... of the Fauna Europaea results, the Biodiversity ... tools to prepare data papers for all 56 major ...
(Date:9/17/2014)... developed, very cleverly, some lessons on how to create ... Joseph Shaw, director of the Optical Technology Center at ... at the nanoscale, we,ll discover them." , Some of ... during a conference called "The Nature of Light: Light ... the University of Arizona College of Optical Sciences. The ...
(Date:9/17/2014)... New research from the Copenhagen Centre for Social Evolution ... evolutionary theory that birth weight and -length can partially ... disorders such as autism and schizophrenia later in life. ... births, and subsequent hospital diagnoses for up to 30 ... factors. The study is published today in the ...
Breaking Biology News(10 mins):Contributions on Fauna Europaea: Data papers as innovative model on expert involvement 2Nature's designs inspire research into new light-based technologies 2Nature's designs inspire research into new light-based technologies 3Size at birth affects risk of adolescent mental health disorders 2Size at birth affects risk of adolescent mental health disorders 3
... -- Traditionally addressed through hand-weeding and/or herbicide application, controlling ... production. Increased labor costs have made hand-weeding prohibitive as ... may be effective and less expensive, non-target herbicide loss ... the environment. To address the economic and environmental impact ...
... -- Associate Professor Paul Blowers of the University of ... 2011 Arizona Professor of the Year, recognized for commitment ... Advancement of Teaching and the Council for Advancement and ... announced the award on Nov. 17 in Washington, D.C. ...
... CAMBRIDGE, Mass. -- Significant progress has been ... psychiatric disease. Recent studies have identified common genetic ... significant risk. One example of a rare cause ... (DISC1) gene, first identified in a large Scottish ...
Cached Biology News:Biodegradable mulches successfully control weeds in container-grown arborvitae 2Arizona Engineering associate professor earns national recognition 2Picower: Schizophrenia gene associated with psychiatric disorders and brain development 2Picower: Schizophrenia gene associated with psychiatric disorders and brain development 3
... The MEGAscript RNAi Kit is ... large mass amounts of ready-to-use ... non-mammalian systems. The MEGAscript RNAi ... patented high-yield transcription technol-ogy and ...
UGT1A6 (D-20)...
... With its patented technology, the Lone Wolf ... any Normally Open proportional valve available on ... operation, provides repeatable high-speed performance, and ensures ... , Enhances system control and patient ...
Normal Hamster Serum...
Biology Products: