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:6/23/2016)... (PRWEB) , ... June 23, 2016 , ... ... Plate® YM (Yeast and Mold) microbial test has received AOAC Research Institute approval ... of microbial tests introduced last year,” stated Bob Salter, Vice President of Regulatory ...
(Date:6/23/2016)... ... , ... Supplyframe, the Industry Network for electronics hardware design ... Located in Pasadena, Calif., the Design Lab’s mission is to bring together inventors ... and brought to market. , The Design Lab is Supplyframe’s physical representation of ...
(Date:6/23/2016)... 23, 2016  Blueprint Bio, a company dedicated to ... medical community, has closed its Series A funding round, ... "We have received a commitment from Forentis ... need to meet our current goals," stated Matthew ... runway to complete validation on the current projects in ...
(Date:6/23/2016)... ... June 23, 2016 , ... ... regulatory and technical consulting, provides a free webinar on Performing Quality ... 13, 2016 at 12pm CT at no charge. , Incomplete investigations are still ...
Breaking Biology Technology:
(Date:6/9/2016)... 9, 2016 Paris Police ... video security solution to ensure the safety of people and ... during the major tournament Teleste, an international technology ... services, announced today that its video security solution will be ... back up public safety across the country. The system roll-out ...
(Date:6/2/2016)... LONDON , June 2, 2016 ... has awarded the 44 million US Dollar project, ... Security Embossed Vehicle Plates including Personalization, Enrolment, and IT Infrastructure ... world leader in the production and implementation of Identity Management ... in January, however Decatur was selected ...
(Date:6/1/2016)... 1, 2016 Favorable Government Initiatives ... and Criminal Identification to Boost Global Biometrics System Market ... TechSci Research report, " Global Biometrics Market By ... and Opportunities, 2011 - 2021", the global biometrics market ... on account of growing security concerns across various end ...
Breaking Biology News(10 mins):