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Transcription profiling of human tumor cell line vs human reference RNA to investigate small sample amplification technologies

ABSTRACT: This sample is part of a study that compares small sample amplification technologies. The analysis looks at differential gene expression when compared to one round of T7 amplification. A tumor cell line was used in comparison to a human reference RNA in this study.

ORGANISM(S): Homo sapiens

SUBMITTER: Chin-Yi Chu

PROVIDER: E-GEOD-2723 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Small Sample Amplification Technologies

Project description:This sample is part of a study that compares small sample amplification technologies. The analysis looks at differential gene expression when compared to one round of T7 amplification. A tumor cell line was used in comparison to a human reference RNA in this study. Keywords = amplification Keywords = small sample Keywords = Affymetrix Keywords: other

2005-06-03 | GSE2723 | GEO

Microarray Based Comparison of two Amplification Methods For Nanogram Amounts of Total RNA

Project description:Two T7 based methods One round of Amplification (Affymetrix) and Two round of Amplification were compared to two Ribo-SPIA based systems, RiboSPIA and pico Ribo SPIA systems. Data for One Round of amplification , Two round of amplification and RiboSPIA are listed here. All Hybridisation were performed using Affymetrix Mouse 430-2 gene chips. Data were all scaled to 500 and scaling factor average was 3.6 +/- 0.9, background intensities 46 +/- 5 , noise 2.9 +/- 0.6, rawQ 1.5 +/- 0.2 Keywords: other

2004-05-28 | GSE1435 | GEO

Microarray Based Comparison of three Amplification Methods For Nanogram Amounts of Total RNA

Project description:Two T7 based methods One round of Amplification (Affymetrix) and Two round of Amplification were compared to two Ribo-SPIA based systems, RiboSPIA and pico Ribo SPIA systems. Data for Pico-RiboSPIA are listed here. All Hybridisation were performed using Affymetrix Mouse 430-2 gene chips. Data were all scaled to 500. For 12 chips performed with pico Ribo SPIA the scaling factor average was 2.0 +/- 0.3, background intensities 74.4 +/- 12.7 , noise 3.9 +/- 0.6, rawQ 2.5 +/- 0.3 Keywords: ordered

2004-12-21 | GSE2019 | GEO

Transcription profiling of mouse samples subject to two different amplification methods for nanogram amounts of total rna to compare amplification methods

2008-06-11 | E-GEOD-1435 | biostudies-arrayexpress

Effect of Low RNA Sample Amount on Amplification Fidelity

Project description:The effect of low RNA sample amount on amplification fidelity was investigated. Keywords: other

2008-02-13 | GSE10436 | GEO

Transcription profiling of Bos taurus immature oocytes and 8-cell embryos to assess the impact of the loss of sample representation caused by the use of global linear amplification (T7-IVT) during microarray hybridization sample preparation

Project description:Assessing the impact of the loss of sample representation caused by the use of global linear amplification (T7-IVT) during microarray hybridization sample preparation.

2009-03-25 | E-MEXP-2099 | biostudies-arrayexpress

Transcription profiling of mouse samples to compare three amplification methods for nanogram amounts of total RNA

Project description:Two T7 based methods One round of Amplification (Affymetrix) and Two round of Amplification were compared to two Ribo-SPIA based systems, RiboSPIA and pico Ribo SPIA systems. Data for Pico-RiboSPIA are listed here. All Hybridisation were performed using Affymetrix Mouse 430-2 gene chips. Data were all scaled to 500. For 12 chips performed with pico Ribo SPIA the scaling factor average was 2.0 +/- 0.3, background intensities 74.4 +/- 12.7 , noise 3.9 +/- 0.6, rawQ 2.5 +/- 0.3

2008-06-11 | E-GEOD-2019 | biostudies-arrayexpress

Trypanosoma cruzi mini-exon based mRNA amplification technique

Project description:We have developed a method of mRNA amplification based on T7 polymerase, where T7 promoter is attached to a oligonucleotide containing the mini-exon sequence, which will hybridize with cDNA molecules containing the anti-ME sequence

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Optimization and evaluation of T7 based RNA linear amplification protocols for cDNA microarray analysis

Project description:Abstract: BACKGROUND: T7 based linear amplification of RNA is used to obtain sufficient antisense RNA for microarray expression profiling. We optimized and systematically evaluated the fidelity and reproducibility of different amplification protocols using total RNA obtained from primary human breast carcinomas and high-density cDNA microarrays. RESULTS: Using an optimized protocol, the average correlation coefficient of gene expression of 11,123 cDNA clones between amplified and unamplified samples is 0.82 (0.85 when a virtual array was created using repeatedly amplified samples to minimize experimental variation). Less than 4% of genes show changes in expression level by 2-fold or greater after amplification compared to unamplified samples. Most changes due to amplification are not systematic both within one tumor sample and between different tumors. Amplification appears to dampen the variation of gene expression for some genes when compared to unamplified poly(A)+ RNA. The reproducibility between repeatedly amplified samples is 0.97 when performed on the same day, but drops to 0.90 when performed weeks apart. The fidelity and reproducibility of amplification is not affected by decreasing the amount of input total RNA in the 0.3-3 micrograms range. Adding template-switching primer, DNA ligase, or column purification of double-stranded cDNA does not improve the fidelity of amplification. The correlation coefficient between amplified and unamplified samples is higher when total RNA is used as template for both experimental and reference RNA amplification. CONCLUSION: T7 based linear amplification reproducibly generates amplified RNA that closely approximates original sample for gene expression profiling using cDNA microarrays. This SuperSeries is composed of the SubSeries listed below.

2006-03-07 | GSE4349 | GEO

Effect of the amount of input total RNA on T7 amplification

Project description:Zhao et al. Amplification Table 9 This experiment was designed to evaluate the effect of the amount of input total RNA on the fidelity, reproducibility, and yield of T7 based RNA linear amplification. BC2 total RNA was amplified using the Jeffrey lab protocol. Different amounts of T7 primer were used according to the quantity of input total RNA. Multiple amplifications were done for each quantity of input total RNA to minimize experimental variations. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set

2005-11-04 | GSE3557 | GEO

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