Publications (PDF files for download)
Publications [R1] to [R3] (section at the top)
The New Product Line “ExpressArt RNAready kits” for RNA isolation is
based on the technology developed by Prof. Rolf Jaggi, Bern University,
Switzerland. Results with this technology are presented here.
Publications [1] to [30]
Results obtained with ExpressArt mRNA amplification kits and other ExpressArt reagents.
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[R3] Antonov, Popovici et al., 2010 (0.5 MB)
Molecular risk assessment of BIG 1-98 participants by expression profiling using RNA from archival tissue. BMC Cancer 2010, 10:37.
The purpose of the work reported here is to test reliable molecular profiles using routinely processed formalin-fixed paraffin-embedded (FFPE)
tissues from participants of the clinical trial BIG 1-98 with a median follow-up of 60 months.
RNAs from fresh frozen (FF) and FFPE tumor samples were isolated with AmpTec’s RNAready and FFPE RNAready kits, respectively. RNAs from FF and FFPE
tumor samples of 82 patients were used for quality control, and independent FFPE tissues of 342 postmenopausal participants of BIG 1-98 with ER-positive
cancer were analyzed by measuring prospectively selected genes.
Pearson correlation coefficients between FF- and FFPE-derived scores were at least 0.94 and high concordance was observed between molecular scores and
immunohistochemical data.
Scores representing the endocrine responsiveness and proliferation status of breast cancers were developed from gene expression analyses based on RNA derived
from FFPE tissues. The validation of the molecular scores with tumor samples of participants of the BIG 1-98 trial demonstrates that such scores can serve as
independent prognostic factors to estimate disease free survival (DFS) in postmenopausal patients with estrogen receptor positive breast cancer.
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[R2] Oberli, Popovici et al., 2008 (1.18 MB)
Expression profiling with RNA from formalin-fixed, paraffin-embedded material. BMC Medical Genomics 2008, 1:9.
A simple procedure with a novel demodification/decrosslinking step was established for RNA isolation from FFPE material of diagnostic tumor samples.
The RNA is suitable for quantitative gene expression measurements by RT-QPCR, and QPCR data were compared for FFPE RNAs obtained with the novel procedure,
or with commercial FFPE RNA isolation kits. The new procedure resulted in consistently lower Ct values, combined with less variability. In comparisons with
intact RNAs isolated from the same tumors (from parallel, fresh-frozen samples), correlation coefficients between intact and RNA from FFPE material were 0.83
to 0.97.
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[R1] Schobesberger et al., 2008 (0.8 MB)
Gene expression variation between distinct areas of breast cancer measured from paraffin-embedded tissue cores. BMC Cancer 2008, 8:343.
Optimized protocols were developed for RNA isolation from histologically distinct areas. RNA prepared from FFPE tissue cores is suitable for gene expression
measurement by quantitative PCR. Distinct molecular scores could be determined from different cores of the same tumor specimen.
Gene expression values were used to calculate scores representing the proliferation status (PRO), the estrogen receptor status and the HER2 status.
The PRO scores measured from entire sections were similar to PRO scores determined from invasive ductal carcinoma (IDC) tissue cores. Scores determined from
normal tissue cores consistently revealed lower PRO scores than cores derived from IDC or ductal carcinoma in situ (DCIS) of the same block or from different blocks of the same patient.
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[31] Englert et al., 2009 (0.5 MB)
Amplification of mRNA without 3’ bias from formalin-fixed paraffin-embedded (FFPE) breast cancer tissues and multiplex expression analysis
on flow-through microarrays. Cancer Res 69(24 Suppl):Abstract nr 3052.
Poster presented at the Thirty-Second Annual CTRC-AACR San Antonio Breast Cancer Symposium. Dec 10-13, 2009.
Total RNA extracted from FFPE samples was subjected to two rounds of amplification with the ExpressArt TRinucleotide kit (AmpTec), which preferentially
primes cDNA synthesis near the 3'-ends of mRNAs (rather than at poly-A sites) and selects for mRNAs against ribosomal RNAs. Biotin-labeled anti-sense RNA (aRNA) was
hybridized to flow-through microarrays on the Ziplex Automated Workstation (Xceed Molecular). Performance was assessed by the analysis of defined titration mixtures
prepared from mRNA from breast cancer and colon cancer FFPE tissue blocks.
Similar signal intensities were observed for 3'-biased probes and probes several hundred bases from the 3'end, confirming that there is no 3' bias in the amplification.
This demonstrated the feasibility of amplifying and quantifying sequences at any position within transcripts in degraded mRNA from FFPE samples. Expression differences of less
than two-fold may be conveniently analyzed with hundreds of probes on the Ziplex Workstation.
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[30] Sharbel et al., 2010 (1.5 MB)
Apomictic and Sexual Ovules of Boechera Display Heterochronic Global Gene Expression Patterns. Plant Cell 22: 655–671.
We have compared the transcriptomic profiles of microdissected live ovules at four developmental stages between a diploid sexual and diploid apomictic Boechera.
We sequenced >2 million SuperSAGE tags and identified (1) heterochronic tags (n=595) that demonstrated significantly different patterns of expression between sexual and
apomictic ovules across all developmental stages, (2) stage-specific tags (n=577) that were found in a single developmental stage and differentially expressed between the sexual
and apomictic ovules, and (3) sex-specific (n=237) and apomixis-specific (n=1106) tags that were found in all four developmental stages, but in only one reproductive mode.
For each accession, 20 ovules per developmental stage were collected in one tube (with two technical replicates for each stage), frozen directly in liquid nitrogen, and stored
at -80°C. RNA was isolated using the PicoPure RNA isolation kit (Arcturus Bioscience).
To prevent degradation of the minute amounts of RNA during the isolation procedure and to enhance the binding efficiency of the RNA on the purification columns, the lysis buffer
was supplemented with 1% (v/v) of NucleoGuard stock solution and 2 mL of N-Carrier (AmpTec).
For the required amounts of dscDNA for the SuperSAGE method, the RNA had to be amplified. Linear mRNA amplification was achieved using the ExpressArt mRNA amplification
kit (AmpTec) with several important modifications. As starting material ~4 ng of total RNA was used in two independent reactions per sample to level out any random methodological effects.
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[29] Murphy, Buckley et al., 2009 (0.5 MB)
Global MYCN Transcription Factor Binding Analysis in Neuroblastoma Reveals Association with Distinct E-Box Motifs and Regions of DNA Hypermethylation. PLoS ONE 4(12): e8154.
Combination of ExpressArt mRNA amplification and Nimblegen microarrays: The ExpressArt TR Micro Kit (Cat. No. 6199-A30, AmpTec) was used to synthesise double-stranded cDNA from 3 ug total RNA,
which was subsequently used to generate amplified amino-allyl antisense RNA (aRNA). The aRNA was coupled to Cy3 reactive dye and 4 µg of Cy3-aRNA was hybridised to the Homo sapiens 4x72K Gene
Expression Array from Roche NimbleGen.
Neuroblastoma, a cancer derived from precursor cells of the sympathetic nervous system, is a major cause of childhood cancer related deaths. The single most important prognostic indicator
of poor clinical outcome in this disease is genomic amplification of MYCN, a member of a family of oncogenic transcription factors.
We applied MYCN chromatin immunoprecipitation to microarrays (ChIP-chip) using MYCN amplified/nonamplified cell lines as well as a conditional knockdown cell line to determine
the distribution of MYCN binding sites within all annotated promoter regions.
Resulting effects on DNA methylation status were correlated with gene expression levels.
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[28] Melkus, Rolletschek et al., 2009 (0.86 MB)
The Metabolic Role of the Legume Endosperm: A Non-invasive Imaging Study. Plant Physiol. 151: 1139–1154.
Although essential for normal seed development in the legumes, the metabolic role of the endosperm remains uncertain. We designed non invasive nuclear magnetic resonance (NMR) tools for the in vivo study on key metabolites in the transient liquid endosperm of intact pea seeds. Expression of mRNA sequences encoding amino acid and sucrose transporters was up-regulated earlier in the endosperm than in the embryo, and this activity led to the accumulation of soluble metabolites in the endosperm vacuole. Microdissection and RT-PCR: Frozen seeds were cut into 10 µm sections and mounted onto PET-membrane slides (Carl Zeiss MicroImaging GmbH). Sections were subjected to laser-assisted microdissection using a PALMfi Laser-Microbeam (Bernried, Germany) device. Small tissue samples were collected using laser pressure catapulting and larger ones by a hand-operated micro-needle. Poly(A)-mRNA was extracted directly using the Dynabeads mRNA DIRECT kit (Dynal Biotech), using the manufacturer’s protocol. Linear amplification and cDNA synthesis was carried out from ~1ng mRNA using the ExpressArt mRNA amplification Nano kit (AmpTec GmbH), according to the manufacturer’s protocol. PCR reactions were based on a template of 20 ng cDNA.
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[27] Sharbel et al., 2009 (1.85 MB)
Molecular signatures of apomictic and sexual ovules in the Boechera holboellii complex. Plant J. 58: 870–882.
The switch to apomixis (the animal analogue is parthenogenesis) is based on de-regulation of developmental pathways originally leading to sexual seed formation.
Individual live ovules were collected, using sterile glass needles. Ten ovules per tube and two
samples per accession were collected in this way, and frozen directly in liquid nitrogen and stored at -80°C. RNA from micro-dissected ovule material was isolated using a PicoPure RNA isolation kit with several modifications. The standard lysis buffer was supplemented with 1% v/v NucleoGuard stock solution and 2 µl N-Carrier
(AmpTec, http://www.amp-tec.com). Linear mRNA amplification was achieved using an ExpressArt mRNA amplification Pico kit (AmpTec) with several modifications. Approximately 1 ng of total RNA was used per sample. The resulting RNA after the first amplification round was purified using RNeasy MinElute columns (Qiagen), followed by a second and third amplification round. Subsequently, 5 µg of DNA-free amplified mRNA was converted into double-stranded cDNA using a 5’-biotinylated primer, and subsequently used for SuperSAGE generation.
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[26] Pietschmann, Beetz et al., 2009 (0.9 MB)
Toll-like receptor expression and function in subsets of human gammadelta T lymphocytes. Scandinavian Journal of Immunology 70: 245-255.
By microarray analysis, we have identified a range of genes differentially regulated in freshly isolated gammadelta T cells by TCR versus TCR plus TLR3 stimulation. RNA was isolated with NucleoSpin RNA-Kit II (Macherey & Nagel) from cell sorter purified peripheral blood gammadelta T cells directly after isolation or after stimulation as described above. For comparison of freshly sorted Vdelta 1 and Vdelta 2 gammadelta T cells, 1 µg total RNA was used for reverse transcription. For comparison of resting and differentially stimulated cells from one donor, 0.5 µg RNA from each sample was amplified using the ExpressArt mRNA Amplification Kit, Micro version (AmpTec, Hamburg, Germany) according to the manufacturer's instructions. About 0.1 µg of the amplified RNA was used for reverse transcription and real time PCR was performed for some of the most interesting differentially expressed genes.
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[25] Nam et al., 2009 (1.0 MB)
Metatranscriptome analysis of lactic acid bacteria during kimchi fermentation with genome-probing microarrays. Int. J. Food Microbiol. 130: 140-146.
We constructed genome probing microarrays (GPM) that are specific to 39 lactic acid bacteria (LAB) in an effort to monitor microbial diversity and biological activity during the fermentation of kimchi, a traditional Korean vegetable product known to contain various health-promoting and immunity-boosting factors.
For RNA extraction, 5 mL aliquots of kimchi soup were pelleted by centrifugation at 5400 g for 10 min. The pellets were then lysed in 1 mL of Trizol reagent (Invitrogen Life Technologies) and 0.4 mL of zirconia-silica beads (0.1 mm diameter; Roth, Karlsruhe, Germany) in a Mini-BeadbeaterTM (Biospec Products, Bartlesville, USA). Labelled cDNAs were prepared from 5 µg RNA samples using the ExpressArt Bacterial mRNA amplification kit.
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[24] Carlucci et al., 2009 (0.3 MB)
A 57-gene expression signature in B-cell chronic lymphocytic leukemia. Biomedicine & Pharmacotherapy 63: 663-671.
The aim of this study is to investigate the gene expression profile, by a specific chip for microarray analysis, in B-CLL lymphocytes with regard to factors involved in apoptosis cascade, signal transduction, purine metabolism enzymes, interleukin expression, enzymes involved in the responses to oxidative stress. We found relevant results in a set of 19 of the 57 genes considered.
Dynabeads were used to enrich for CD19+ cells. About 5 µg total RNA per sample, were amplified and labelled with the ExpressArt Aminoallyl mRNA Amplification Kit, Micro Version. Cy-labelled RNAs were hybridised on a specific custom-chip with 57 gene targets.
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[23] Thoudam et al., 2008 (40 KB)
Differential gene expression profile of stomach and oral cancer in high risk region of India. Genomic Med. (2008) 2:376
RNA extracted from ten tumor tissues (five with oral squamous cell carcinoma and five with gastric adenocarcinoma) was amplified using ExpressArt AminoAllyl mRNA amplification
Kit and labelled with NHS-activated Cy-3 dye. Normal tissue obtained from a site distant to tumour was used as control. The labelled probes were hybridized with Human 40K OciChip Array
containing 20,106-probes.
Gene Ontology revealed that genes involved in regulation of actin cytoskeleton, MAPK and Wnt signalling pathways were differentially expressed in both cancers. Genes involved in Neuroactive ligand-receptor interaction and leukocyte transendothelial migration were
differentially expressed in gastric adenocarcinoma exclusively. Genes of TGF beta signalling pathway were significantly downregulated in gastric adenocarcinoma while in oral cancer
genes of T-cell-receptor and Toll like receptor pathway were significantly upregulated.
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[22] Knight et al., 2008 (425 KB)
TEAD1 and c-Cbl are novel prostate basal cell markers that correlate with poor clinical outcome in prostate cancer. Br. J. Cancer 99:1849-1858
Immunohistochemistry of clinical prostate cancer samples was combined with laser microdissection to isolate 200 to 1,000 luminal and basal cells. Isolated RNAs were amplified with the ExpressArt TR Nano kit, followed by hybridisation to genome-wide cDNA arrays. Differential expression data were confirmed by RT-PCR and immunostaining.
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[21] Morris et al., 2008 (521 KB)
Epstein–Barr virus-encoded LMP1 induces a hyperproliferative and inflammatory gene expression programme in cultured keratinocytes. J. Gen. Virol. 89: 2806-2820
Affymetrix microarray data were obtained with microgram quantities of RNA by using standard Affymetrix technology, or with nanogram amounts of RNA and amplification with the ExpressArt Nano kit. Differential expression data were generated with three different analysis methods. Microarray data were extensively validated by qRT-PCR, ELISA, immunofluorescence and Western blots.
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[20] Schain et al., 2008 (392 KB)
Evidence for a pathophysiological role of cysteinyl leukotrienes in classical Hodgkin lymphoma. Int. J. Cancer: 12: 2285–2293.
RNA was isolated from fresh-frozen clinical biopsies, from Hodgkin Lymphoma cell lines, LMP1-transfected GC B-cells and EBV-infected KMH2 cells. 150–200 H-RS cells and 200–300 nonmalignant cells were isolated by laser microdissection from Hodgkin Lymphoma tumors and amplified with the ExpressArt Nano kit. Microarray data were confirmed by qPCR and immunohistochemistry.
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[19] Vockerodt et al., 2008 (425 KB)
The Epstein–Barr virus oncoprotein, latent membrane protein-1, reprograms germinal centre B cells towards a Hodgkin’s Reed–Sternberg-like phenotype. J. Pathol. 216: 83-92.
RNA from FACS-sorted transfected GC B cells was amplified with the ExpressArt mRNA Amplification kit. Microarray data were evaluated by qRT-PCR, immunohistochemistry, immunofluorescence and Western blots.
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[18] Baumforth et al., 2008 (1.78 MB)
Expression of the Epstein-Barr Virus-Encoded Epstein-Barr Virus Nuclear Antigen 1 in Hodgkin’s Lymphoma Cells Mediates Up-Regulation of CCL20 and the Migration of Regulatory T Cells. Am. J. Pathol. 173:195-204.
Microdissections were performed on cryosections using the PALM laser microdissector. For microarray analysis (Affymetrix HG-U133 Plus 2.0 GeneChips), RNA was extracted from 150-200 Hodgkin Reed-Sternberg cells or from GCs. RNA was amplified in two rounds with the ExpressArt TRinucleotide mRNA amplification kit.
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[17] Campean et al., 2008 (570 KB)
Angiopoietin 1 and 2 gene and protein expression is differentially regulated in acute anti-thy 1.1. Am J Physiol Renal Physiol 294: F1174-F1184.
Laser microdissection (MMI AG, Switzerland), and use of ExpressArt N-Carrier ("Pico RNA Care") for RNA isolation, followed by analysis on Affymetrix RAE230A Microarray GeneChip.
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[16] Su et al., 2008 (424 KB)
Lambda light chain revision in the human intestinal IgA response. J Immunol 181:1264–1271.
Laser microdissection (P.A.L.M. Microlaser Technologies), and use of ExpressArt N-Carrier ("Pico RNA Care") for RNA isolation, followed by analysis with RT-PCR.
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[15] Hu et al., 2008 (256 KB)
Exon-level expression profiling: a comprehensive transcriptome analysis for oral fluids. Clin. Chem. 54(5): 824-832.
Conclusion: "It is feasible to use [very small] samples containing fragmented RNAs to conduct high-resolution expression profiling with coverage of the entire transcriptome and to validate multiple targets from limited amounts."
Goal was the analysis of amplified salivary RNA with Affymetrix Exon Arrays with the challenges of small sample size (< 5 ng total human RNA) and severe RNA degradation. The standard Affymetrix protocol with rRNA removal with magnetic beads (requires >100 ng of intact RNA) and subsequent RNA amplification by random priming (no selectivity against rRNAs, and requires >100 ng RNA) was impossible.
AmpTec's universal mRNA-specific linear amplification strategy (ExpressArt TRinucleotide mRNA amplification kit) has overcome both limitations. It provides selection against rRNAs, thus rRNA removal is obsolete, and two amplification rounds are possible to provide the necessary amounts of amplified mRNAs for Exon arrays. Array results were validated by qPCR data and an expression phenotype (male/female) was correctly identified.
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[14] Yanai, Baugh et al., 2008 (345 KB)
Pairing of competitive and topologically distinct regulatory modules enhances patterned gene expression. Mol. Syst. Biol. 4:163.
Study of the embryonic development of Caenorhabditis elegans is hampered by the fact that a complete organism provides only ~ 200 picograms of total RNA. Whole-genome microarray
analysis necessitates extensive mRNA amplification. Although a sophisticated version of the standard, Eberwine-based technology has been developed [Baugh et al. (2001), Quantitative analysis
of mRNA amplification by in vitro transcription. Nucleic Acids Res. 29:E29.], a minimum of 10 pooled embryos was still required.
A modification of ExpressArt technology [now available as ExpressArt Pico mRNA amplification kit, #8399-A12] has enabled this large-scale study with sub-nanogram RNA amounts. The RNA from 10 embryos could be safely split in
half to provide a back-up for further analyses.
"We used RNAi and time series, whole-genome microarray analyses to systematically perturb and characterize components of a Caenorhabditis elegans lineage-specific transcriptional regulatory network. These data are supported by selected reporter gene analyses and comprehensive yeast one-hybrid and promoter sequence analyses."
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[13] Cummings et al., 2008 (286 KB)
Sexual and social stimuli elicit rapid and contrasting genomic responses. Proc. R. Soc. B 275: 393-402.
An initial evaluation of ExpressArt technology was performed [Duftner et al., 2008] and here, ExpressArt is applied in a differential gene expression study.
"Here we examine preference behaviour of 58 female swordtails, Xiphophorus nigrensis, in four different social environments (attractive and unattractive males, females only, non-attractive males only and asocial conditions) followed by neural gene expression profiling. We used a brain-specific cDNA microarray to identify patterns of genomic response and candidate genes, followed by quantitative PCR (qPCR) examination of gene expression with variation in behaviour. Our microarray results revealed patterns of genomic response differing more between classes of social stimuli than between presence versus absence of stimuli. ..... The identification of stimulus- and behaviour-specific responses opens an exciting window into the molecular pathways associated with social behaviour and mechanisms that underlie sexual selection."
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[12] Duftner et al., 2008 (804 KB)
Efficacy of RNA amplification is dependent on sequence characteristics: implications for gene expression profiling using a cDNA microarray. Genomics 91: 108-117.
Detailed study on potential bias introduced by mRNA amplification with the ExpressArt Nano mRNA amplification kit, provides also a brief overview of the ExpressArt technology. ExpressArt
technology was subsequently used in a differential gene expression study [Cummings et al., 2008].
Obtained yields: "Assuming that 1–5% of total RNA correspond to poly(A)+ RNA, the first round of amplification yielded an approximately 1000- to 4000-fold increase, while the second
round yielded an approximately 200-fold increase of poly(A)+ equivalents. These amplification factors correlate well with the expected values given by the manufacturer of the amplification
kit for 500 ng input material in both the first and the second round of amplification."
"We applied a T7 polymerase-based technique [ExpressArt] to amplify RNA from two tissues of a cichlid fish and compared expression levels of unamplified and amplified RNA on a cDNA microarray. Amplification bias was generally minor and comprised features that were lost (1.3%) or gained (2.5%) through amplification and features that were scored as regulated before but unregulated after amplification (4.2%) or vice versa (19.5%). We examined 10 sequence-specific properties and found that GC content, folding energy, hairpin length and number, and lengths of poly(A) and poly(T) stretches significantly affected RNA amplification. We conclude that, if RNA amplification is used in gene expression studies, preceding experiments controlling for amplification bias should be performed."
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[11] Zhang et al., 2008 (0.97 MB)
We describe a simple and highly effective means for global identification of genes that are expressed within specific cell types within complex tissues. It involves transgenic
expression of nuclear-targeted green fluorescent protein in a cell-type-specific manner. The fluorescent nuclei are then purified from homogenates by fluorescence-activated sorting, and
the RNAs employed as targets for microarray hybridization. We demonstrate the validity of the approach through the identification of 12 genes that are selectively expressed in phloem.
Two consecutive rounds of amplification were done on 5 µl samples (approximately 2–20 ng RNA), using ExpressArt mRNA amplification kits (Nano plus Version; Artus GmbH) according to the manufacturer’s instructions. Aminoallyl-modified UTP was incorporated into the amplified antisense RNA produced during the second round of in vitro transcription.
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[10] Birgersdotter et al., 2007 (725 KB)
Three-dimensional culturing of the Hodgkin lymphoma cell-line L1236 induces a HL tissue-like gene expression pattern. Leukemia and Lymphoma 48: 2042-2053.
"To overcome some limitations of in vitro established cell-line tumor models for Hodgkin lymphoma (HL), we explored whether culturing in a three-dimensional (3D) matrix could improve the quality of the model..... The gene expression profile of the 3D-cultured HL derived cell-line L1236 was compared with that of suspension-cultured (2D) L1236, as well as to the gene expression profile found in HL tumor samples.
To validate our results we also included a gene expression data set of laser captured Hodgkin- Reed-Sternberg (H-RS) cells..... Between 150 and 200 H-RS cells or nonmalignant cells were isolated.... Following RNA extraction, three rounds of linear T7-based mRNA amplification were performed using the ExpressArt system....
The gene expression profiles were analyzed using Affymetrix technology. We found that the 3D culture affected gene expression of an HL derived cell-line, inducing a more tumor-related expression profile."
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[9] Bose et al., 2007 (646 KB)
Down-regulation of ATM protein in HRS cells of nodular sclerosis Hodgkin's lymphoma in children occurs in the absence of ATM gene inactivation. J Pathol 213:329-336.
Microdissections were performed on cryosections using the Leica or PALM laser microdissector. For microarray analysis (Affymetrix HG-U133 Plus 2.0 GeneChips), RNA was extracted from 200-250 Hodgkin Reed-Sternberg cells and amplified in two rounds with the ExpressArt Nano mRNA amplification kit.
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[8] Eickhoff et al., 2007 (548 KB)
Host cell responses to Chlamydia pneumoniae in gamma-interferon-induced persistence overlap those of productive infection and are linked to genes involved in apoptosis, cell cycle and metabolism. Infection and Immunity 75: 2853-2863.
RNA was isolated from cultured HeLa cells, to compare expression profiles of persistently infected cells with those of mock-infected cells. The ExpressArt Micro mRNA amplification kit was used for subsequent analysis with Affymetrix HG-U133A GeneChips. A high level of technical reproducibility was found, and a set of nineteen differentially expressed genes were selected for qPCR analysis. Microarray data were confirmed for seventeen genes.
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[7] Elvidge, 2006 (1.1 MB)
Microarray expression technology: from start to finish. Pharmocogenomics 7: 123-134.
Technology Report: An overview of methods and platforms is presented, including ExpressArt mRNA amplification with its unique ability to perform up to 3 amplification rounds and using sub-nanogram RNA quantities, combined with adaptations for degraded RNAs. |
[6] Wetzel et al., 2006 (325 KB)
Bone morphogenetic protein-7 expression and activity in the human adult normal kidney is predominantly localized to the distal nephron. Kidney Int. 70: 717-723.
Laser microdissection (MMI AG, Switzerland), and use of ExpressArt N-Carrier ("Pico RNA Care") for RNA isolation, followed by qPCR.
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[5] McLucas et al., 2006 (187 KB)
Global gene expression analysis of the effects of Vinblastine on endothelial cells, when eluted from a thermo-responsive polymer. J. Biomed. Mater. Res. A: 246-253.
Use of ExpressArt mRNA amplification kit (Nano version) with subsequent hybridisation to human 40K OciChip.
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[4] Reis et al., 2006 (322 MB)
Isolation of mouse neuritic mRNAs. J Mol Histol. 37: 79-86.
Laser microdissection and use of ExpressArt mRNA amplification kit (Pico version) to visualize isolated RNA.
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[3] Gomez-Mancilla et al., 2005 (0.47 MB)
Central Nervous System Drug Development: An Integrative Biomarker Approach toward Individualized Medicine. NeuroRx. 2: 683-695.
This review represents an effort to illustrate the integration of state of the art and emerging technologies in drug development supporting the path of individualized medicine.
Citation: "Today, ExpressArt mRNA amplification technology based on TRinucleotide primers allows the complete amplification of all mRNA fragments in severely degraded RNA samples and works very well with extremely low limits of input RNA amounts (picogram range)."
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[2] Okuducu et al., 2005 (1.48 MB)
Cellular retinoic acid-binding protein 2 is down-regulated in prostate cancer. Int. J. Oncol. 27: 1273-1282.
Laser microdissection, subsequent use of ExpressArt mRNA amplification kit (2 amplification rounds), followed by cDNA array hybridisation.
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[1] Zeng et al., 2005 (343 KB)
Transcription factor Gfi1 regulates self-renewal and engraftment of hematopoietic stem cells. EMBO J. 23: 4116-4125.
ExpressArt Carrier ("Pico RNA Care") for RNA isolation from FACS-sorted cells.
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