Navigation Links
Use of Transcriptor Reverse Transcriptase,,,in Microarray Analysis

nitor gene expression in both host and pathogen [1]. However, the system must be optimised for pathogen gene expression since the T. gondii RNA can be more than an order of magnitude less concentrated compared with the host cell RNA. Over the past few years, technological advances have made the method much more reliable and robust.

Curiously, the standard protocols for generating the labelled cDNA target from the experimental RNA, arguably the most critical step in the procedure, remain virtually unchanged. The vast majority of investigators use either a direct incorporation of deoxynucleotides with covalently attached fluorophores or the incorporation of amino-allyl deoxynucleotides with subsequent fluorophore conjugation. In both methods, the efficiency of the target labelling reaction and quality of the labelled cDNA product depend almost entirely on the reverse transcription enzyme used.

We employ an amino-allyl labelling reaction to generate and compare microarray data from different commercially available reverse transcriptases.

Materials and Methods

Parasite and RNA preparation

Parasites (RH strain) were cultured according to a standard protocol. NIH 3T3 cells were used as host cells, and the parasites were harvested by centrifugation just prior to monolayer lysis. The cells were suspended in TRI reagent (Sigma). RNA was extracted according to the manufacturers protocol, except that the RNA was not ethanol precipitated following incubation with 2-propanol, but was added directly to an RNeasy Midi column (Qiagen). Binding of the sample was followed directly with the recommended wash and two extra washes. The RNA was eluted with a total of 450 l RNase-free water. This was followed by a standard
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Transcriptor Reverse Transcriptase
2. Increase the Power and Sensitivity for Your cDNA Synthesis with the New Transcriptor Reverse Transcriptase
3. Properties of Reverse Transcriptases
4. Reverse transcriptases suitable for one-step RT-PCRb
5. Reverse transcriptases suitable for two-step RT-PCRd
6. C. therm. Polymerase for Reverse Transcription in Two-Step RT-PCR
7. Recombinant HIV Reverse Transcriptase
8. Novel Bonding Chemistry Imparts Enhanced Polar Selectivity to TSK-GEL ODS-100V Reversed Phase Columns
9. New Microarray Labeling Kit for Preparing cDNA Probes
10. Stratagenes Human cDNA Microarrays: Perform Gene Expression Profiling and Discover New Genes
11. An Optimal DNA Microarray Substrate for the Identification of Fetal and Somatic Liver Stem Cells of the Rat
Post Your Comments:
(Date:1/15/2014)... 2014  Bill Jacobs Automotive, a group of car dealerships headquartered ... the Heartland Blood Center and offering free oil change coupons ... place Saturday, Jan. 18 at the Bill Jacobs Cadillac and ... (Photo: http://photos.prnewswire.com/prnh/20140115/MN46637 )  The Bill Jacobs ...
(Date:1/15/2014)...   Niagara Thermal Products is proud to announce ... LLC ("B3") as the critical thermal component supplier in ... Army Corps of Engineers.  B3 has developed the only ... and destroys VOCs and other chemical pollutants, allergens and ...
(Date:1/15/2014)... 2014 Cynvenio, a cancer diagnostics company ... genomic analysis of tumor cells in the bloodstream, today ... cancer decision support application powered by CollabRx, Inc. ... genomic test, Therapy Finder will provide breast cancer patients ...
(Date:1/15/2014)... Jan. 15, 2014 ­ RedBrick Health , a ... technology, today announces that EmblemHealth , the ... company, is now providing the RedBrick Compass health assessment, ... to all of its members. EmblemHealth is among the ...
Breaking Biology Technology:Join the Bill Jacobs Auto blood drive and receive a free oil change 2Niagara Thermal Products LLC Selected as Critical Supplier for Buffalo BioBlower Air Purification Systems 2Cynvenio 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 3EmblemHealth Selects RedBrick Health to Power its Next-Generation Health Engagement and Behavior Change System 2EmblemHealth Selects RedBrick Health to Power its Next-Generation Health Engagement and Behavior Change System 3
... Anne T. Ferguson, Ph.D., ... , INTRODUCTION , This application ... from Molecular Devices to study the performance of the LIVE/DEAD ... fluorescent dyes, calcein AM (cal AM) and ethidium homodimer (EthD-1), to ...
... , A Sensitive, Robust, Homogenous,and Rapid Assay , ... method , Assays a broad range of tyrosine kinases ... in a kit and in bulk packaging , , ... activity can be accelerated using High Efficiency Fluorescence Polarization ...
... David Handfield, Martin Boissonneault, Esther Tremblay, Benoit Houle and Luc Mnard. ... , Recent advances in ... in both the number of compounds and potential drug targets that ... assay systems has helped to meet the demand for increased throughput. ...
Cached Biology Technology:LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43) 2LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43) 3LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43) 4LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43) 5TKXtra DataFile: Homogenous tyrosine kinase detection using High Efficiency Fluorescence Polarization 2TKXtra DataFile: Homogenous tyrosine kinase detection using High Efficiency Fluorescence Polarization 3Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets. 2Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets. 3Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets. 4Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets. 5
(Date:4/17/2014)... RSV infection , Respiratory syncytial virus (RSV) ... infections, generating life-threating illness in very young and ... limited. RSV enters host cells through the fusion ... bundle. Small interfering peptides that prevent bundle formation ... are highly susceptible to degradation. In this issue ...
(Date:4/17/2014)... Professor Marc Andr Gauthier and Professor Luca Razzari ... each been awarded large grants from the John ... for Innovation (CFI) for the acquisition of state-of-the-art ... be added matching grants from the Ministre de ... et de la Technologie (MESRST). These new laboratories ...
(Date:4/17/2014)... cells in bone marrow need to produce hydrogen sulfide ... according to a new study from the Center for ... Dentistry of USC. , Professor Songtao Shi, principal investigator ... produced by the cells governs the flow of calcium ... signals that results in osteogenesis, or the creation of ...
Breaking Biology News(10 mins):JCI online ahead of print table of contents for April 17, 2014 2JCI online ahead of print table of contents for April 17, 2014 3JCI online ahead of print table of contents for April 17, 2014 4JCI online ahead of print table of contents for April 17, 2014 5JCI online ahead of print table of contents for April 17, 2014 6New state-of-the-art biotech and nanotech equipment for INRS 2
... is available in German . , Scientists of ... investigated another navigational skill of desert ants. These ants are ... sun compass along with a step counter and visible landmarks ... research team from Jena recently discovered that these ants also ...
... of Biological Sciences (AIBS) has selected Meredith Niles, a ... Richards, a graduate student at the University of Maryland, ... the University of Rhode Island, to receive the 2010 ... "AIBS is committed to fostering a productive dialogue between ...
... Americans recognize things need to change in the grocery ... is included in their food and on the packages. ... the right food choices to avoid obesity, but will readily ... new survey by FoodMinds. "In light of all the ...
Cached Biology News:Smelling the scenery in stereo 2AIBS names emerging public policy leaders 2AIBS names emerging public policy leaders 3Americans want Uncle Sam's help putting healthy foods on their dinner table 2
... 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: