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:1/14/2014)... AUSTIN, Texas , Jan. 14, 2014 ... analyze and research medicinal plants and therapeutic derivatives ... industry, health professionals, and researchers about the challenges ... (Logo: http://photos.prnewswire.com/prnh/20100430/DC95601LOGO ) The ...
(Date:1/14/2014)... January 14, 2014 iLab Solutions, the leader ... the new Director of Product Strategy. In this role, Michelle ... iLab sub-teams to guide in the development of iLab products. ... the maximum possible benefit to the scientific community by offering ...
(Date:1/14/2014)... Jan. 14, 2014  RXi Pharmaceuticals Corporation (OTCQX: RXII), ... commercializing innovative therapies addressing major unmet medical needs ... the Notice of Allowance from the United States ... RNAi compounds (sd-rxRNA®), for the treatment of fibrosis. ...
(Date:1/14/2014)... ®, provider of RapidResponse ®, a cloud-based supply chain ... S&OP ) service, is proud to be a gold sponsor ... at the Hilton San Diego Bayfront, January 30 – 31, ... , Director of Supply Chain at Amgen, as she presents ...
Breaking Biology Technology:World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 2World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 3World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 4World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 5World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 6World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 7World's Largest Group of Medicinal Plant Researchers Endorses ABC-AHP-NCNPR Botanical Adulterants Program 8iLab Solutions Announces Michelle Detwiler as the New Director of Product Strategy 2RXi Pharmaceuticals Receives US Notice of Allowance for a Key Patent Relating to its Self-Delivering Technology with sd-rxRNAs targeting CTGF, including RXI-109, for the Treatment of Fibrotic Disorders 2RXi Pharmaceuticals Receives US Notice of Allowance for a Key Patent Relating to its Self-Delivering Technology with sd-rxRNAs targeting CTGF, including RXI-109, for the Treatment of Fibrotic Disorders 3Event Alert: Kinaxis Customer to Present at the Biomanufacturing Summit "Supply Chain: Improving Network Effectiveness" 2
... , which has promised Milwaukee that it can provide citywide ... reprieve by the Milwaukee Common Council after falling ... give Midwest Fiber until June 7 to hook up the ... The company was to have the area hooked up by ...
... this, about half of all Americans required to do so are scurrying ... in Sunday's Arizona Republic . For a lot of folks like ... back by two days this year, was indeed upon us. And we'd ... of tax laws changed this year. , ,The realization that Tax Day ...
... stuff of legends. Sit in any presentation by one ... PowerPoint slide you'll likely be convinced that the methodology ... itself, and eventually bring about world peace in our ... unique and special. When they are questioned about the ...
Cached Biology Technology:Turbo Tax Online can teach new Web concepts a business lesson 2Turbo Tax Online can teach new Web concepts a business lesson 3With tech implementations, it's NOT the methodology, stupid 2With tech implementations, it's NOT the methodology, stupid 3
(Date:4/23/2014)... and German . ... the difference between sexes depends on one single element of the ... the two sexual chromosomes are X and Y, whereas women have ... the morphological and physiological differences between males and females. , But ... ago, the X and Y were identical, until the Y started ...
(Date:4/23/2014)... is available in French and Spanish ... 23 April 2014Today, the CEC released its Conservation Assessment ... Approach to Conservation, which identifies 29 priority conservation areas ... includes 11 different protected areas in the states of ... diverse arid and semi-arid habitats inhabited by rare and ...
(Date:4/23/2014)... People who have had a stroke, often suffer motor ... a study conducted in Taiwan, that will be published ... but is currently freely available on-line as an unedited ... when one group of stroke victims had their own ... brain and a similar group did not, those who ...
Breaking Biology News(10 mins):Male or female? 2Male or female? 3Conservation priorities released for several protected areas along US-Mexico border 2Conservation priorities released for several protected areas along US-Mexico border 3Autologous stem cell therapy improves motor function in chronic stroke victims 2
... YORK (Feb. 28, 2013) The Democratic Republic of ... Okapi Faunal Reserve (OFR), has declined by 37 percent in ... according to wildlife surveys by WCS and DRC officials. ... DRC continues unabated, the species could be nearly extinguished from ...
... source of great controversyparticularly in Europebut acts of vandalism ... the health and ecological impacts of those crops hard ... for use in GM crop studies could serve as ... crop biotechnology, according to an article published in ...
... 2,778 bees while retracing the muddy steps of a scientist ... than a century ago. Occasionally stung, but considering herself ... guided her field work, the Montana State University ecologist and ... ." "It,s exciting," Burkle said as the Feb. 28 ...
Cached Biology News:Elephants are vanishing from DRC's best-run reserve 2Elephants are vanishing from DRC's best-run reserve 3Elephants are vanishing from DRC's best-run reserve 4Fighting GM crop vandalism with a government-protected research site 2Old records, new bees result in 'Science' paper for MSU ecologist 2Old records, new bees result in 'Science' paper for MSU ecologist 3
... Novagen's HT96 Isothermal Block is an ... hold HT96 plates and to provide efficient ... plate. This new design of the HT96 ... offers an enhanced product design allowing for ...
Umbilical cord blood neutrophils, For immunohistochemistry (IHC)...
...
Adjustable tilt angle 0 to 20 Adjustable speed of 3-25 RPM Load capacity: 2.7 kg (6 lbs.) Ambient temperature range 4C to 65C Low voltage power is supplied by wall mount transformer...
Biology Products: