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:1/15/2014)... Calif. (PRWEB) January 15, 2014 Cynvenio, ... and management through the genomic analysis of tumor cells ... Therapy Finder™, a web-based cancer decision support application powered ... option with Cynvenio’s ClearID™ genomic test, Therapy Finder will ...
(Date:1/15/2014)... , January 15, 2014 A study ... races on the Formula 1 track could help to tackle ... Applied Technologies (MAT), Stowhealth (a GP surgery based in Stowmarket) ... healthcare provider Simplyhealth. Telemetry technology, which is inspired ...
(Date:1/15/2014)... January 15, 2014 DTS Language Services, ... Online Web Portal for Life Science organizations who need document ... the subject matter of their documents in advance with a ... reduce time-to-delivery of translations, often a critical factor in clinical ...
(Date:1/15/2014)... TAIPEI , Jan. 15, 2014 TaiGen Biotechnology ... an exclusive agreement with R-Pharm, a leading Russian pharmaceutical ... in the Russian Federation , ... Commonwealth Independent States (CIS). Nemonoxacin is a novel antibiotic for ...
Breaking Biology Technology:Cynvenio Offers New Breast Cancer Therapy Finder Application for its ClearID Breast Cancer Genomic Test 2Cynvenio Offers New Breast Cancer Therapy Finder Application for its ClearID Breast Cancer Genomic Test 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 2Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 4DTS Improves Efficiency for Life Science Document Translations 2TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 2TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 3TaiGen Biotechnology Signed Exclusive License Agreement with R-Pharm for Nemonoxacin (Taigexyn(R)) 4
... Genzyme Corp. said on Wednesday it has agreed ... Wisconsin-Madison biotech spinoff, for $600 million in cash. , ,Bone ... more than $2 billion and almost 7,000 employees worldwide, plan ... companies recently reported healthy quarterly profits and revenues. Bone Care ...
... a new copy machine for $57,000. , ,The Z-Corp ... typical office copier: It makes three-dimensional models of small structures ... , ,Using white polymer powder and colored inks, the printer ... a time. Z-Corp is headquartered in Burlington, Massachusetts. , , ...
... can submit executive summaries of their businesses for angel ... May 9. , ,The Wisconsin Angel Network ... investor networks with entrepreneurs based on the investors' market ... a deal-flow pipeline, has been designed to be a ...
Cached Biology Technology:Massachusetts firm to buy UW spinoff Bone Care for $600 million 2Massachusetts firm to buy UW spinoff Bone Care for $600 million 3Start-ups, angels can connect on new state Web site 2Start-ups, angels can connect on new state Web site 3
(Date:4/17/2014)... honored Clemson professor Rajendra Singh Thursday as a ... and expand solar deployment in the residential, commercial ... Banks Professor of Electrical and Computer Engineering and ... considered a local hero leading the charge across ... in solar power and driving policy changes at ...
(Date:4/17/2014)... honeybees in Europe, Asia and the United States are ... be impacting native honeybee populations at this time, according ... include including Nosema microsporidia and Varroa ... resilient to these invasive pests, which suggests to us ... Asia and the United States currently are not necessary ...
(Date:4/17/2014)... Researchers in France have hit on a novel ... receive the correct and most effective treatment possible., ... the western and developing world. If left untreated, ... to renal failure and other complications. In many ... problem. Clearly a more effective pathological approach to ...
Breaking Biology News(10 mins):White House honors Clemson professor as 'Champion of Change' for solar deployment 2East African honeybees are safe from invasive pests… for now 2East African honeybees are safe from invasive pests… for now 3
... throughout the world and ensuring safe water supplies would do ... any other possible measure, according to an analysis released today ... water and sanitation reduces poverty in three ways. ... entrepreneurs; , Significant savings are achieved in the ...
... Researchers have discovered a mechanism for the rapid growth seen ... The tumors, which are made up of proliferating blood ... descent, with girls more frequently afflicted than boys. The ... blood-red lump on the head or facethen grow rapidly in ...
... Missouri Botanical Garden in St. Louis operates one of ... and the second largest in the western hemisphere. With ... a gigantic new aroid species from Ecuador, the ... has reached a milestone of six million specimens. ...
Cached Biology News:Providing toilets, safe water is top route to reducing world poverty: UN University 2Providing toilets, safe water is top route to reducing world poverty: UN University 3Providing toilets, safe water is top route to reducing world poverty: UN University 4Providing toilets, safe water is top route to reducing world poverty: UN University 5Researchers identify Achilles heel of common childhood tumor 2Missouri Botanical Garden mounts milestone 6 millionth herbarium specimen 2Missouri Botanical Garden mounts milestone 6 millionth herbarium specimen 3
... Automatic grid, subgrid and spot finding overcome tedious spot, ... template assignment , ... options to optimize, ... Characterize more precisely through grouping by cells, grids, ...
... unique human proteins on a 1x3 ... a cross section of all cellular ... high-quality results. Additional control proteins and ... validity and reproducibility., ,UNIchip protein microarrays ...
...
... The benchmark for analytical performance ... ease-of-use and the ultimate in ... Varian 810-MS features include: ... optics minimize,interferences, providing reliable, routine, ...
Biology Products: