Navigation Links
Amplification of Mouse cDNAs for Microarrays Using the Eppendorf ,,, MasterTaq Kit

After printing, the microarrays were allowed to dry completely at ambient conditions. The slides were pretreated with 2x SSPE, 0.2% SDS at 55C. The cDNA probe was lyophilized and redissolved in ~32 l Microarray Hybridization Buffer Version 2 (APBiotech, UK). The solution was added to the microarray and a coverslip was applied. Hybridization was allowed to proceed for 1418 hours at 42C. The microarray was washed with 1X SSC/0.2% SDS for 5 minutes at 45C followed by two 5-minute washes with 0.1X SSC/0.2% SDS at room temperature. The microarray was rinsed briefly with water, dried with nitrogen, and scanned using a Molecular Dynamics Generation III scanner.

Results and Discussions

For the analysis of gene expression in eukaryotes, expressed sequence tag (EST) data represent an invaluable resource for gene identification. EST's are single pass partial cDNA sequences. In general, cDNA clones are selected to represent as many unique transcripts as possible. One such method to group transcripts is the Unigene clustering of data sets which attempts to identify unique human transcripts within EST data (http://www.ncbi.nlm.nih.gov/UniGene). In amplifying EST collections for the production of microarrays, the yield of each PCR amplicon is a very important consideration. The optimal concentration of DNA for successful microarray experimentation is typically 200 g/ml. Hybridizations using non-covalent attachment chemistries (e.g. polylysine, aminosilane) generally work poorly with DNA spotted at concentrations lower than this. When amplifying thousands of clones, the main objective is to minimize the total number of reactions needed to generate sufficient material for printing. A high yield o
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9

Related biology technology :

1. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
2. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
3. Greater Amplification Specificity with New Hot Start PCR Enzyme
4. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
5. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
6. Improve Amplification Specificity with Hot Start PCR Enzyme
7. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
8. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
9. Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler
10. Amplification of Epstein-Barr Virus Exon C with Difficult Primers Using Eppendorf Mastercycler gradient
11. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
Post Your Comments:
(Date:9/19/2014)... 19, 2014 BioTech News Coverage: ... and results of studies and trials.  Companies in focus ... (NYSE: LLY ), Gilead Sciences Inc. (NASDAQ: ... CELG ) and Arena Pharmaceuticals Inc. (NASDAQ: ARNA) ... prodrug therapeutics for the treatment of cancer, announced that ...
(Date:9/19/2014)... September 19, 2014 Cereal grain ... industries will gather October 5–8, 2014, for the ... Island Convention Center in Providence, Rhode Island. The ... work of food scientists, chemists, microbiologists, nutritionists, and ... food production management. To date, 1,000 attendees have ...
(Date:9/19/2014)... , September 19, 2014 , ... European Society for Paediatric Endocrinology (ESPE) Meeting  ... and medical research in the field of growth  , ... Darmstadt, Germany , today announced the ... for 2014. The awards were announced during a Satellite ...
(Date:9/18/2014)... 18, 2014 Bipartisan passage this week ... designed to stimulate development and commercialization of new technologies ... by leaders of SPIE, the international society for ... Manufacturing and Innovation (RAMI) Act, now moves to the ... said they are optimistic that it will win passage ...
Breaking Biology Technology:Biotech Companies Pushing Sector Higher With Recent Therapeutic Advancements & Patent Approvals - Company Wins Summary Judgment For Prodrug Patent 2Biotech Companies Pushing Sector Higher With Recent Therapeutic Advancements & Patent Approvals - Company Wins Summary Judgment For Prodrug Patent 3Biotech Companies Pushing Sector Higher With Recent Therapeutic Advancements & Patent Approvals - Company Wins Summary Judgment For Prodrug Patent 4Biotech Companies Pushing Sector Higher With Recent Therapeutic Advancements & Patent Approvals - Company Wins Summary Judgment For Prodrug Patent 5Biotech Companies Pushing Sector Higher With Recent Therapeutic Advancements & Patent Approvals - Company Wins Summary Judgment For Prodrug Patent 6Biotech Companies Pushing Sector Higher With Recent Therapeutic Advancements & Patent Approvals - Company Wins Summary Judgment For Prodrug Patent 7Latest Research on Cereal Grain Quality, Health Benefits and Food Production to be Presented During the AACC International Annual Meeting 2Latest Research on Cereal Grain Quality, Health Benefits and Food Production to be Presented During the AACC International Annual Meeting 3EMD Serono Awards Grant for Growth Innovation (GGI) for the First Time 2EMD Serono Awards Grant for Growth Innovation (GGI) for the First Time 3House passage of high-tech manufacturing bill is move toward stronger economy, say SPIE leaders 2House passage of high-tech manufacturing bill is move toward stronger economy, say SPIE leaders 3
... tracking the number and dimension of particles or droplets as they ... are specific and possibly unique, changes that are of critical importance ... provide you with the knowledge, information and best practices you need ... ...
... of a new system to study crystallization processes for ... name,Crystalline. , The Crystalline has ... to learn as much as possible about a drug ... "Avantium is proud to launch its next system ...
... Institute have discovered a new method for controlling ... potentially one step closer to realizing the mass ... sheet of carbon, was discovered in 2004 and ... silicon as the fundamental building blocks of nanoelectronics. ...
Cached Biology Technology:Maximize Data Analysis with METTLER TOLEDO FBRM 2Avantium Launches the Crystalline, a New Parallel System for Crystallization Research 2Light-speed nanotech: Controlling the nature of graphene 2
(Date:9/18/2014)... a world-leading provider of scientific, technical and medical ... Management Society (ARMS), recognized the academic research achievements ... Young Researcher of the Year Awards. , The ... Convention Centre and in conjunction with the ARMS ... Australia,s and New Zealand,s scientific community. Guest of ...
(Date:9/18/2014)... is probably the ultimate form of camouflage: you don,t ... this strategy is not as uncommon as you might ... County, USA, explains that the larval life stages of ... part of the anatomy that most creatures cannot make ... have to shield each individual eye unit with an ...
(Date:9/17/2014)... to reproductive technologies, the justice and well-being of ... to identify key issues, articulate their values and ... most defensible ways forward. But what are the ... The Hastings Center and the Presidential Commission for ... to publish a series of essays to highlight ...
Breaking Biology News(10 mins):Elsevier and Australasian Research Management Society award scientists at Scopus ceremony 2Transparent larvae hide opaque eyes behind reflections 2Why bioethics literacy matters 2Why bioethics literacy matters 3
... This release is available in German . ... blood cells and the functioning of the nervous system. Unable to ... via animal proteins. So far it has been known that on ... organelles, so-called lysosomes. From there, the vitamin enters the cell interior ...
... CITY, MOTwo opposing teams battle it out to regulate ... activators, keeps DNA open to enzymes that transcribe DNA ... twisting DNA into an inaccessible coil around histone proteins, ... by enzymes that elongate an RNA strand. Both ...
... scientists have, for the first, time created a type ... functional, bio-compatible nanoscale wires into engineered human tissues. ... in Nature Materials , a multi-institutional research team ... Professor of Chemistry at Harvard and Daniel Kohane, a ...
Cached Biology News:Vitamin B12 deficiency: Tracing the causes 2Controlling gene expression: How chromatin remodelers block a histone pass 2Controlling gene expression: How chromatin remodelers block a histone pass 3Merging the biological and the electronic 2Merging the biological and the electronic 3
... Kit is designed for concentration and ... samples. Proteomics sample analysis is often ... impurities such as buffers, salts and ... interfere with electrophoretic separations or enzymatic ...
... offers maleimide activated carrier proteins to ... Imject Maleimide Activated Carrier Proteins are ... preactivated with a heterobifunctional cross-linker (Sulfo-SMCC). ... sulfhydryl-reactive carrier protein. Thus allowing ...
... kit allows for rapid, convenient, and efficient ... bind/wash/elute process utilizes a spin column to ... for concentration of proteins which were electro-eluted ... strong ion exchangers, offering high binding capacity ...
... polystyrene with frosted labeling surfaces, volume ... counting. 100% integrity tested. Linkage: This ... number, created to easily match Cornings ... yet, please order under the old ...
Biology Products: