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:
(Date:8/3/2015)... , Aug. 3, 2015  ArroGen Group, ... its new Fingerprint Molecular Identification™ (FMID) technology Aug. ... conference in Sacramento, Calif. ... science, will enable law enforcement agents, district attorneys ... of criminal suspects. By analyzing chemical ...
(Date:8/3/2015)... WESTWOOD, Mass. , Aug. 3, 2015  Merck ... company for innovative and top-quality high-tech products in the ... together with the US-based company Nano-C, their development and ... field of organic photovoltaics (OPV). Merck KGaA, Darmstadt, ... participate in the growing OPV sector. Novel ...
(Date:8/3/2015)... 2015   Intrexon Corporation (NYSE: XON ... second quarter 2015 financial results after the market closes ... will host a conference call at 5:30 PM ET ... update. The conference call may be accessed ... asking to join the "Intrexon Corporation Call."  Participants may ...
(Date:7/31/2015)... , July 31, 2015 In today,s ... that help them drive positive health outcomes, improve ... source of patient care in their communities. At ... tradeshow and conference, AmerisourceBergen announced a new set ... Services Administration Organization and help independent pharmacies endure ...
Breaking Biology Technology:ArroGen Previews New Fingerprint Molecular Identification Technology 2Merck KGaA, Darmstadt, Germany, and Nano-C Present Enhanced Materials for Organic Photovoltaics 2Merck KGaA, Darmstadt, Germany, and Nano-C Present Enhanced Materials for Organic Photovoltaics 3Merck KGaA, Darmstadt, Germany, and Nano-C Present Enhanced Materials for Organic Photovoltaics 4Intrexon to Announce First Half and Second Quarter 2015 Financial Results and Host Conference Call on August 10th 2AmerisourceBergen Unveils Suite of Enhanced Capabilities for Independent Pharmacies at ThoughtSpot 2015 2AmerisourceBergen Unveils Suite of Enhanced Capabilities for Independent Pharmacies at ThoughtSpot 2015 3
... VALENCIA, Calif., June 16 MannKind,Corporation (Nasdaq: MNKD ), ... announced today that it,will present at upcoming conferences., ... 23, 2008 at 11:45 AM (Eastern ... Island, Massachusetts, -- 2nd Annual Jefferies Healthcare Conference ...
... FRANCISCO, June 16 The Endocrine Society, an,international association ... for security analysts. The call,originated from ENDO 08, the ... in San Francisco. The Society has over 14,000 members ... report on the latest,advances in endocrine research and clinical ...
... 16 Celator Pharmaceuticals today,announced that John Bennett, ... the BIO Business Forum during the Biotechnology Industry,Organization ... will,provide a corporate overview and an update on ... on Wednesday, June 18,2008 at 3:30 PDT in ...
Cached Biology Technology:'Breakthroughs Ahead,' Endocrine Society Tells Security Analysts 2
(Date:7/9/2015)... 07, 2015 Research ... of the "Biometrics for Banking; Market & ... report to their offering. The adoption ... this growth and the forecast is that by ... for companies involved in delivering biometric systems to ...
(Date:7/8/2015)... , July 8, 2015  Trovagene, Inc. (NASDAQ: ... diagnostics, today announced the launch of a study ... Precision Cancer Monitoring℠ (PCM) technology for predicting response ... or a combination of the novel immunotherapy agents ... a PD-1 inhibitor. The 50-patient study will be ...
(Date:7/2/2015)... June 25, 2015 ... addition of the "Natural Language Processing Market ... IVR, OCR, Pattern & Image Recognition) - Worldwide ... The key vendors occupying the market ... Corporation, Microsoft Corporation, NetBase Solutions, SAS Institute Inc., ...
Breaking Biology News(10 mins):Global Banking Biometrics Market 2015-2020 - Market & Technology Analysis, Adoption Strategies and Forecasts 2Global Banking Biometrics Market 2015-2020 - Market & Technology Analysis, Adoption Strategies and Forecasts 3Trovagene Initiates Study to Monitor Response to Immunotherapy in Melanoma Patients Using its Precision Cancer Monitoring Platform 2Trovagene Initiates Study to Monitor Response to Immunotherapy in Melanoma Patients Using its Precision Cancer Monitoring Platform 3Trovagene Initiates Study to Monitor Response to Immunotherapy in Melanoma Patients Using its Precision Cancer Monitoring Platform 4Natural Language Processing Market by Type (Rule-Based, Statistical, and Hybrid), Technologies (Recognition, IVR, OCR, Pattern & Image Recognition) - Worldwide Forecast to 2020 2
... asthma has been shown to help reduce the formation ... is implicated in the development of Alzheimer,s disease, according ... The researchers published their findings, "Pharmacologic Blockade of 5-Lipoxygenase,Improves ... in the American Journal of Pathology . ...
... The long tendrils of this woody vine, or liana, ... But kudzu may be no match for the lianas of ... studies show that lianas are overgrowing trees in every instance. ... forest ecosystems. Tropical forests contain more than half of ...
... The Pancreatic Cancer Action Network and the American Association ... and E. Scott Seeley, M.D., Ph.D., the 2011 Pancreatic ... each totaling $600,000 over five years, will be formally ... April 2-6. "Because of the poor prognoses and ...
Cached Biology News:Asthma drug could help control or treat Alzheimer's disease 2Stranglers of the tropics -- and beyond 2Stranglers of the tropics -- and beyond 3Pancreatic Cancer Action Network-AACR Pathway to Leadership Grants awarded to early-career investigators 2Pancreatic Cancer Action Network-AACR Pathway to Leadership Grants awarded to early-career investigators 3Pancreatic Cancer Action Network-AACR Pathway to Leadership Grants awarded to early-career investigators 4
Endomorphin-2. Cross reactivity: Endomorphin-1 3%, Met-enkephalin <0.03%, Leu-enkephalin <0.01% and beta-Endorphin <0.01%....
... Error-free cloning of BAC (100+kb), fosmid ... Greater insert stability than any othe BAC ... for high yields and easy recombinant DNA ... or transcription-free (pSMART VC) vectors for the ...
... is used in conjunction with ... of problematic and/or GC-rich templates ... with 10X PCRX Amplification Buffer ... magnesium concentration broader annealing temperature ...
MAGP-2 (C-19)...
Biology Products: