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cdna microarray facility  (AECOM International Development)

 
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    AECOM International Development cdna microarray facility
    Complementary-template reverse-transcription (CT-RT) of single-stranded DNA primers reverse-transcribed from FFPE-RNA. ( a ) RNA extracted from FFPE tissue is reverse-transcribed, the mRNA/DNA duplex is filtered on an YM-50 column and the DNA is single-stranded with RNase-H and column purified. The 5′-NB-Oligo-dA (24) -cT7-3′ (complementary to the T7 promoter) is annealed to the <t>FFPE-cDNA</t> primers. ( b ) Total RNA from universal human reference (UHR, Stratagene) is amplified using the Sense-Amp cRNA amplification kit from Genisphere to provide RNA with the same orientation as messenger RNA . ( c ) Single-stranded DNA primers are hybridized to their sense-RNA template between 70 and 42°C for 90 min. The hybridized products are reverse-transcribed by a process described as CT-RT. The restored FFPE-cDNAs are doubled stranded and transcribed in vitro using T7 polymerase. (See Supplementary Data for technical description of points 1 through 6.)
    Cdna Microarray Facility, supplied by AECOM International Development, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/dna+microarray+chip+method/pmc01976468-129-11-19?v=AECOM+International+Development
    Average 90 stars, based on 1 article reviews
    cdna microarray facility - by Bioz Stars, 2026-08
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    1) Product Images from "Molecular restoration of archived transcriptional profiles by complementary-template reverse-transcription (CT-RT)"

    Article Title: Molecular restoration of archived transcriptional profiles by complementary-template reverse-transcription (CT-RT)

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkm510

    Complementary-template reverse-transcription (CT-RT) of single-stranded DNA primers reverse-transcribed from FFPE-RNA. ( a ) RNA extracted from FFPE tissue is reverse-transcribed, the mRNA/DNA duplex is filtered on an YM-50 column and the DNA is single-stranded with RNase-H and column purified. The 5′-NB-Oligo-dA (24) -cT7-3′ (complementary to the T7 promoter) is annealed to the FFPE-cDNA primers. ( b ) Total RNA from universal human reference (UHR, Stratagene) is amplified using the Sense-Amp cRNA amplification kit from Genisphere to provide RNA with the same orientation as messenger RNA . ( c ) Single-stranded DNA primers are hybridized to their sense-RNA template between 70 and 42°C for 90 min. The hybridized products are reverse-transcribed by a process described as CT-RT. The restored FFPE-cDNAs are doubled stranded and transcribed in vitro using T7 polymerase. (See Supplementary Data for technical description of points 1 through 6.)
    Figure Legend Snippet: Complementary-template reverse-transcription (CT-RT) of single-stranded DNA primers reverse-transcribed from FFPE-RNA. ( a ) RNA extracted from FFPE tissue is reverse-transcribed, the mRNA/DNA duplex is filtered on an YM-50 column and the DNA is single-stranded with RNase-H and column purified. The 5′-NB-Oligo-dA (24) -cT7-3′ (complementary to the T7 promoter) is annealed to the FFPE-cDNA primers. ( b ) Total RNA from universal human reference (UHR, Stratagene) is amplified using the Sense-Amp cRNA amplification kit from Genisphere to provide RNA with the same orientation as messenger RNA . ( c ) Single-stranded DNA primers are hybridized to their sense-RNA template between 70 and 42°C for 90 min. The hybridized products are reverse-transcribed by a process described as CT-RT. The restored FFPE-cDNAs are doubled stranded and transcribed in vitro using T7 polymerase. (See Supplementary Data for technical description of points 1 through 6.)

    Techniques Used: Reverse Transcription, Purification, Amplification, In Vitro

    Experimental procedure utilized for the analysis of 10-year-old matched frozen and formalin-fixed paraffin embedded breast cancer samples. ( a ) Five micrograms of RNA extracted from the 10-year-old frozen portion of the sample, is reverse-transcribed and the cDNA is double stranded (dsDNA), in four individual reactions. The dsDNA of three reactions undergoes IVT-amplification (MessageAmpII, Ambion), which gives rise to complementary RNA (cRNA) for cDNA microarray analyses. The dsDNA of one reaction is used for PCR experiments. ( b ) Five micrograms of RNA extracted from the 10-year-old FFPE portion of the sample underwent the exact same process. ( c ) Single-stranded DNA (ssDNA) obtained by RT of 5 μg of FFPE-RNA is purified and hybridized to the sense-RNA template library. The restored ssDNA is double stranded and purified. Three of the CT-RT reactions undergo IVT-amplification, while the dsDNA of one reaction is used for PCR experiments.
    Figure Legend Snippet: Experimental procedure utilized for the analysis of 10-year-old matched frozen and formalin-fixed paraffin embedded breast cancer samples. ( a ) Five micrograms of RNA extracted from the 10-year-old frozen portion of the sample, is reverse-transcribed and the cDNA is double stranded (dsDNA), in four individual reactions. The dsDNA of three reactions undergoes IVT-amplification (MessageAmpII, Ambion), which gives rise to complementary RNA (cRNA) for cDNA microarray analyses. The dsDNA of one reaction is used for PCR experiments. ( b ) Five micrograms of RNA extracted from the 10-year-old FFPE portion of the sample underwent the exact same process. ( c ) Single-stranded DNA (ssDNA) obtained by RT of 5 μg of FFPE-RNA is purified and hybridized to the sense-RNA template library. The restored ssDNA is double stranded and purified. Three of the CT-RT reactions undergo IVT-amplification, while the dsDNA of one reaction is used for PCR experiments.

    Techniques Used: Formalin-fixed Paraffin-Embedded, Reverse Transcription, Amplification, Microarray, Purification

    Signal intensity and heat-map analysis of the correlation between the log 2 ratios measured by cDNA microarrays. ( a ) Signal intensity of one sample grid in the red channel (Cy5) across all microarrays. Top three panels display the grids obtained from three repeats using cRNA from 10-year-old frozen RNA (Frozen-Amp 1–3). Three mid-panels show the signal of three repeats using cRNA obtained by restoration and IVT-amplification of RNA from 10-year-old FFPE tissue (FFPE-Restored 1–3). Three bottom panels display the signal of three repeats using cRNA obtained by direct IVT-amplification of RNA from 10-year-old FFPE tissue. ( b ) Heat map displaying the log 2 of expression ratios ranging between 0.5 and 2 for 1044 genes detected in frozen tissue on a 28 032 features cDNA microarray and represented in the UHR library. From left to right are displayed the ratios obtained by IVT-amplification of RNA from 10-year-old frozen tissue (Frozen-Amp 1–3), restoration and IVT-amplification of RNA from 10-year-old FFPE tissue (FFPE-Restored 1–3) and direct IVT-amplification of RNA from 10-year-old FFPE tissue (FFPE-Amp 1–3). Each column represents an individual hybridization and each line a different feature. Red and blue represent up-regulated and down-regulated genes, respectively.
    Figure Legend Snippet: Signal intensity and heat-map analysis of the correlation between the log 2 ratios measured by cDNA microarrays. ( a ) Signal intensity of one sample grid in the red channel (Cy5) across all microarrays. Top three panels display the grids obtained from three repeats using cRNA from 10-year-old frozen RNA (Frozen-Amp 1–3). Three mid-panels show the signal of three repeats using cRNA obtained by restoration and IVT-amplification of RNA from 10-year-old FFPE tissue (FFPE-Restored 1–3). Three bottom panels display the signal of three repeats using cRNA obtained by direct IVT-amplification of RNA from 10-year-old FFPE tissue. ( b ) Heat map displaying the log 2 of expression ratios ranging between 0.5 and 2 for 1044 genes detected in frozen tissue on a 28 032 features cDNA microarray and represented in the UHR library. From left to right are displayed the ratios obtained by IVT-amplification of RNA from 10-year-old frozen tissue (Frozen-Amp 1–3), restoration and IVT-amplification of RNA from 10-year-old FFPE tissue (FFPE-Restored 1–3) and direct IVT-amplification of RNA from 10-year-old FFPE tissue (FFPE-Amp 1–3). Each column represents an individual hybridization and each line a different feature. Red and blue represent up-regulated and down-regulated genes, respectively.

    Techniques Used: Amplification, Expressing, Microarray, Hybridization



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    Image Search Results


    The layout of the DNA microarray method module; each detection panel includes 4 detection modules, which can detect two specimens at the same time. Modules 1 and 3 are used to detect mutations in the rpoB gene, and modules 2 and 4 are used to detect mutations in the katG gene and inhA promoter. QC quality control probe; EC external control probe; BC blank control; NC negative control probe; IC internal control probe; WT wild-type.

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    Figure Lengend Snippet: The layout of the DNA microarray method module; each detection panel includes 4 detection modules, which can detect two specimens at the same time. Modules 1 and 3 are used to detect mutations in the rpoB gene, and modules 2 and 4 are used to detect mutations in the katG gene and inhA promoter. QC quality control probe; EC external control probe; BC blank control; NC negative control probe; IC internal control probe; WT wild-type.

    Article Snippet: The CapitalBio DNA microarray chip method is used to qualitatively detect nucleic acids in samples of Mtb isolates from clinical TB patients.

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    The drug susceptibility test was used as a standard method to evaluate the efficacy of the DNA microarray for detecting RIF and INH resistance and MDR-TB.

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    Figure Lengend Snippet: The drug susceptibility test was used as a standard method to evaluate the efficacy of the DNA microarray for detecting RIF and INH resistance and MDR-TB.

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    Comparison of the diagnostic efficacy of the DNA microarray method when sputum smear grades were ≤ 1 + and ≥ 2 +

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    Figure Lengend Snippet: Comparison of the diagnostic efficacy of the DNA microarray method when sputum smear grades were ≤ 1 + and ≥ 2 +

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    Specimen processing procedure: A total of 5,911 sputum smear-positive specimens were collected. After experimental processing, 4148 specimens that were positive with the DNA microarray method and DST were finally included in the study. NTM, nontuberculous mycobacteria ; DST, drug sensitivity test.

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    Pattern diagrams of several common drug-resistant gene mutations detected by the DNA microarray method. The white box is the detection site of the wild-type codon, and the red box is the site of the detected mutant codon. ( a ) rpoB gene Leu511Pro (CTG → CCG); ( b ) rpoB gene Asp516Tyr (GAC → TAC); ( c ) rpoB gene His526Tyr (CAC → TAC); ( d ) rpoB gene Ser531Trp (TCG → TGG); ( e ) rpoB gene Ser531Leu (TCG → TTG); ( f ) katG gene Ser315Thr (AGC → ACC); ( g ) katG gene Ser315Asn (AGC → AAC); ( h ) inhA gene promoter-15 (C → T).

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    Figure Lengend Snippet: Pattern diagrams of several common drug-resistant gene mutations detected by the DNA microarray method. The white box is the detection site of the wild-type codon, and the red box is the site of the detected mutant codon. ( a ) rpoB gene Leu511Pro (CTG → CCG); ( b ) rpoB gene Asp516Tyr (GAC → TAC); ( c ) rpoB gene His526Tyr (CAC → TAC); ( d ) rpoB gene Ser531Trp (TCG → TGG); ( e ) rpoB gene Ser531Leu (TCG → TTG); ( f ) katG gene Ser315Thr (AGC → ACC); ( g ) katG gene Ser315Asn (AGC → AAC); ( h ) inhA gene promoter-15 (C → T).

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    Figure Lengend Snippet: Non-tuberculous species identified from 2013 to 2018 in a tertiary hospital in Beijing, China

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    Specimen types among which NTM were identified in this study

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    Figure Lengend Snippet: Specimen types among which NTM were identified in this study

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