Navigation Links
Microarray Target Purification Kit and,,,the Microarray Target Synthesis Kit (T7)

As the human genome sequence project is completed, the vast amount of genomic information calls for methods that allow gene expression analysis on a genomic scale and speed up detection of significant sequence variations. The most suitable tools available for gene expression analysis are DNA hybridization arrays that allow researchers to examine differences in expression of hundreds or thousands of genes in parallel. That is why DNA arrays, on a variety of platforms such as macroarrays, microarrays or high-density oligonucleotide arrays make their way into different areas of functional genomics research. Arrays provide immobilized gene-specific sequences (probes) on different solid supports (e.g., nylon membranes, glass slides, or silicon/ceramic chips) and are hybridized with labeled targets that are copies of nucleic acids derived from various biological sources.



This requires reliable and efficient tools for preparation and labeling of targets: RNA needs to be extracted from small amounts of precious sample materials, and labeled either directly during cDNA preparation or via a linear amplification (100 to 1000 fold) with T7 polymerase producing labeled cRNA, or via polymerase chain reaction (PCR)-based amplification methods. This step must be highly reproducible and efficient to avoid differences in hybridization caused by biased amplification of target material.

Therefore, Roche Applied Science has developed a range of DNA microarray analysis products for expression profiling, suitable especially for very small amounts of starting material or when highest sensitivity is required. Each product is designed and tested in combination with the others to cover the critical workflow from RNA isolation to preparation of labeled targets. After each step, it is essential that residual impurities or nucleases are removed from the reaction products (single-stranded [ss] or double-stranded [ds] cDNA, cRNA or PCR products). Therefore, we have developed a nucleic acid purification kit that is universally applicable to the whole range of potential samples.

Product Description

cDNA Synthesis System

The well-established cDNA Synthesis System provides a convenient one-tube procedure for the synthesis of ds cDNA, starting even with small amounts of total RNA (120 g). Using the supplied Oligo ([dT24] T7 promoter) primer, cDNA is synthesized containing a T7 RNA polymerase promoter for the subsequent transcription reaction with T7 RNA polymerase.

Microarray Target Purification Kit

The Microarray Target Purification Kit is specifically designed for the purification of a variety of synthesized nucleic acids in the target labeling and amplification process, such as:
  • ss cDNA after 1st strand cDNA synthesis
  • ssDNA after cDNA labeling
  • ds cDNA after 2nd strand cDNA synthesis
  • PCR products
  • cRNA after in-vitro transcription

The kit will remove residual nucleotides, primers, proteins, nucleases and other reaction components which will otherwise interfere in the workflow for highly sensitive microarray hybridization. The process does not require nucleic acid precipitation, phenol/chloroform extractions, or extensive handling of nucleic acids.

Microarray Target Synthesis Kit (T7)

The Microarray Target Synthesis Kit (T7) has been developed as a robust, reliable and highly reproducible tool for the preparation of large amounts of labeled cRNA when starting with only 105 106 cells for target preparation. Major features are:
  • acceptance of low amounts of cDNA (obtained from 1 g total RNA)
  • short reaction time of only 2 3 hours (Figure 2)
  • high yields of cRNA in the range of 100 g
  • compatible to a broad spectrum of hapten or fluorophore-labeled nucleotides
  • cost-effective labeling due to optimized nucleotide concentrations and reaction conditions



Targets, synthesized and labeled with the Microarray Target Synthesis Kit (T7) have been tested on various commercially available DNA arrays. We have found that it is sufficient to use only one labeled nucleotide to get reliable results compared to double-labeling procedures in the majority of experiments.

Very often sample material is available in such small quantities (starting with less than 1,000 cells) that efficient target preparation requires PCR-based amplification methods. Within the Microarray Target Synthesis Kit (PCR), we have optimized a random primer-based PCR amplification procedure which allows the efficient and reproducible amplification of mRNA out of 50 ng total RNA.

The resulting PCR product can then be labeled by different methods, e.g., with the Microarray RNA Target Synthesis Kit (T7) to obtain single stranded cRNA.




'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. New Microarray Labeling Kit for Preparing cDNA Probes
2. Stratagenes Human cDNA Microarrays: Perform Gene Expression Profiling and Discover New Genes
3. An Optimal DNA Microarray Substrate for the Identification of Fetal and Somatic Liver Stem Cells of the Rat
4. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
5. Use of Transcriptor Reverse Transcriptase in Microarray Analysis
6. Increased Sensitivity in Microarray Analysis
7. Analysis of Microarray Data
8. Cancer-Related miRNAs Uncovered by the mirVana miRNA Microarray Platform
9. Is RNA Amplification Necessary for Microarrays?
10. Perform Microarray Analysis with Limited Bacterial RNA
11. Isolate High Quality Total RNA from LCM Samples Suitable for Microarray and qRT-PCR Analysis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/11/2017)... ... 2017 , ... At its national board meeting in North ... the co-founder, CEO and chief research scientist of Minnesota-based Advanced Space Technology and ... ARCS Alumni Hall of Fame . ASTER Labs is a technology development ...
(Date:10/11/2017)... ... October 11, 2017 , ... ComplianceOnlineā€™s Medical Device Summit is back for ... June 2018 in San Francisco, CA. The Summit brings together current and former FDA ... board directors and government officials from around the world to address key issues in ...
(Date:10/11/2017)... and LAGUNA HILLS, Calif. , Oct. 11, ... Research, London (ICR) and University of ... SkylineDx,s prognostic tool to risk-stratify patients with multiple myeloma (MM), ... nine . The University of Leeds ... funded by Myeloma UK, and ICR will perform the testing ...
(Date:10/11/2017)... ... October 11, 2017 , ... Singh Biotechnology today ... designation to SBT-100, its novel anti-STAT3 (Signal Transducer and Activator of Transcription 3) ... able to cross the cell membrane and bind intracellular STAT3 and inhibit its ...
Breaking Biology Technology:
(Date:5/23/2017)... GENOA, Italy , May 23, 2017  Hunova, the first robotic ... and trunk, has been officially launched in Genoa, Italy ... Europe and the USA . The ... launched on the market by the IIT spin-off Movendo Technology thanks to ... view the Multimedia News Release, please click: ...
(Date:5/6/2017)... , May 5, 2017 RAM ... announced a new breakthrough in biometric authentication based ... quantum mechanical properties to perform biometric authentication. These new ... semiconductor material created by Ram Group and its ... entertainment, transportation, supply chains and security. Ram Group ...
(Date:4/18/2017)... Inc., a global expert in SoC-based imaging and computing solutions, has ... features the company,s hybrid codec technology. A demonstration utilizing TeraFaces Ā® ... be showcased during the upcoming Medtec Japan at Tokyo Big Sight ... Las Vegas Convention Center April 24-27. ... Click here for an image of the ...
Breaking Biology News(10 mins):