cdna  (Qiagen)


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    Structured Review

    Qiagen cdna
    Cdna, supplied by Qiagen, used in various techniques. Bioz Stars score: 97/100, based on 478 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna/product/Qiagen
    Average 97 stars, based on 478 article reviews
    Price from $9.99 to $1999.99
    cdna - by Bioz Stars, 2020-02
    97/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: 200 μ L chloroform was added and tubes were shaken vigorously for 15 sec and transferred to 2 mL Eppendorf tubes followed by centrifugation for 15 min at 12000 × g . .. All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions.

    Amplification:

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: As negative control for PCR amplification, reactions with RNA in the absence of reverse transcriptase as well human peripheral blood and mouse embryonic fibroblast cells were included. .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    Synthesized:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: .. All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions. .. Briefly, the volume of RNA was adjusted to 500 ng and mixed with 2 μ L buffer GE and RNase‐free water to 10 μ L. This genomic DNA elimination mix was heated for 5 min at 42°C and immediately placed on ice for 2 min. Nine μ L reverse transcriptase mix (mixed according to manufacturer's instructions) was then added to the RNA mix and incubated at 42°C for 15 min, followed by 5 min at 95°C.

    Article Title: TGR5 potentiates GLP-1 secretion in response to anionic exchange resins
    Article Snippet: .. Quantitative RT-PCR RNA was isolated from cells using TRIreagent (Ambion), after which cDNA was synthesized (Qiagen). .. Quantitative RT-PCR was performed using SYBR green (Roche) in the Lightcycler 480 II (Roche).

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells
    Article Snippet: .. RNA was isolated and analyzed as indicated above. cDNA was synthesized (Qiagen, RT first strand kit) and the PCR array was performed according to the manufactures protocol (Qiagen, Cat.no. ..

    Quantitative RT-PCR:

    Article Title: TGR5 potentiates GLP-1 secretion in response to anionic exchange resins
    Article Snippet: .. Quantitative RT-PCR RNA was isolated from cells using TRIreagent (Ambion), after which cDNA was synthesized (Qiagen). .. Quantitative RT-PCR was performed using SYBR green (Roche) in the Lightcycler 480 II (Roche).

    Article Title: CXCL17 Chemokine-Dependent Mobilization of Memory CXCR8+CD8+ TEM and TRM Cells in the Vaginal Mucosa is Associated with Protection Against Genital Herpes
    Article Snippet: Paragraph title: Quantitative RT-PCR ... Equal concentrations of RNA were used for the reverse transcription reaction to generate cDNA (Qiagen, Valencia, CA).

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA). .. Predesigned PCR primers for the tested genes were purchased from IDT technologies (Coralville, IA Real-time RT-PCR was performed with 200 ng of total RNA and results were analyzed using the 2−ΔΔCt relative quantification method, after normalization to beta-actin.

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. Quantitative gene expression analysis was performed by TaqMan-based QRT-PCR on ABI 7700 (Applied Biosystems, Foster City, CA).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: Paragraph title: Reverse transcriptase-polymerase chain reaction (RT-PCR) and Quantitative PCR (RT-qPCR) ... The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    SYBR Green Assay:

    Article Title: TGR5 potentiates GLP-1 secretion in response to anionic exchange resins
    Article Snippet: Quantitative RT-PCR RNA was isolated from cells using TRIreagent (Ambion), after which cDNA was synthesized (Qiagen). .. Quantitative RT-PCR was performed using SYBR green (Roche) in the Lightcycler 480 II (Roche).

    Article Title: Ovine middle cerebral artery characterization and quantification of ultrastructure and other features: changes with development
    Article Snippet: The mRNA was reverse-transcribed using the Quantitect RT kit, which includes a DNase treatment step before making cDNA (Qiagen). .. Real-time PCR was performed on a LightCycler (model 1.5, Roche, Indianapolis, IN) using the SYBR Green method.

    Article Title: The BH3 mimetic HA14-1 enhances 5-fluorouracil-induced autophagy and type II cell death in oesophageal cancer cells
    Article Snippet: RNA extraction and real-time PCR RNA was extracted, treated with DNase (Ambion, Dublin, Ireland) and cDNA synthesised (Qiagen). .. Each real-time PCR reaction contained 0.5 μ of each primer, MgCl2 (Noxa 4 m, β -actin 3 m), 1 × LightCycler FastStart DNA Master SYBR Green and template cDNA.

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA). .. Briefly, total RNA was isolated from cells and cDNA was prepared using RT2 First Strand kit (SA Biosciences, Frederick, MD) and SYBR green based real-time PCR was carried out using StepOnePlus Real-Time PCR machine (Applied Biosystems, Foster City, CA).

    Article Title: Influence of Aspergillus fumigatus conidia viability on murine pulmonary microRNA and mRNA expression following subchronic inhalation exposure
    Article Snippet: .. RT2 SYBR Green ROX qPCR Mastermix was used for quantification of cDNA (Qiagen, CA). .. Data were normalized by gene expression relative to the levels of glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) for the Th1–Th2 response pathway and beta-glucuronidase ( Gusb ) for the Th17 response pathway.

    Incubation:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions. .. Briefly, the volume of RNA was adjusted to 500 ng and mixed with 2 μ L buffer GE and RNase‐free water to 10 μ L. This genomic DNA elimination mix was heated for 5 min at 42°C and immediately placed on ice for 2 min. Nine μ L reverse transcriptase mix (mixed according to manufacturer's instructions) was then added to the RNA mix and incubated at 42°C for 15 min, followed by 5 min at 95°C.

    Article Title: Bis-anthracycline WP760 abrogates melanoma cell growth by transcription inhibition, p53 activation and IGF1R downregulation
    Article Snippet: 70% confluency were incubated with 100 nM WP760 or DMSO (control) for 24 h. Subsequently, cells were trypsinized, washed with PBS-, lysed with RLT buffer containing β-ME (Qiagen), and stored at −80 °C until RNA isolation. .. Total RNA (3 μg) was reverse-transcribed into cDNA (Qiagen, 330,404) and qPCR was performed using Qiagen reagents (330502) and CFX96 thermocycler (Bio-Rad).

    Expressing:

    Article Title: Bis-anthracycline WP760 abrogates melanoma cell growth by transcription inhibition, p53 activation and IGF1R downregulation
    Article Snippet: PCR array analysis RT2 Profiler PCR Array (Qiagen, PAHS-507Z) was used for gene expression analysis. .. Total RNA (3 μg) was reverse-transcribed into cDNA (Qiagen, 330,404) and qPCR was performed using Qiagen reagents (330502) and CFX96 thermocycler (Bio-Rad).

    Article Title: TGR5 potentiates GLP-1 secretion in response to anionic exchange resins
    Article Snippet: Quantitative RT-PCR RNA was isolated from cells using TRIreagent (Ambion), after which cDNA was synthesized (Qiagen). .. All mRNA expression levels were corrected for expression of the housekeeping gene 36B4 or β2-microglobulin.

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA). .. To investigate the altered expression of chemoresistance-related genes, PCR-based array for drug resistance specific genes (PAHS004Z, Cancer Drug Resistance PCR Array; SA Biosciences, Frederick, MD) was used.

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. Quantitative gene expression analysis was performed by TaqMan-based QRT-PCR on ABI 7700 (Applied Biosystems, Foster City, CA).

    Article Title: Influence of Aspergillus fumigatus conidia viability on murine pulmonary microRNA and mRNA expression following subchronic inhalation exposure
    Article Snippet: Paragraph title: Gene Expression Arrays ... RT2 SYBR Green ROX qPCR Mastermix was used for quantification of cDNA (Qiagen, CA).

    Activated Clotting Time Assay:

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. The TaqMan primers and probe for TMPRSS2 were: forward primer: CAC GGA CTG GAT TTA TCG ACA A; reverse primer: CGT CAA GGA CGA AGA CCA TGT, probe: TGA GGG CAG ACG GCT A; for PSA/KLK3 forward primer: CCC ACT GCA TCA GGA ACA AA, reverse primer: GAG CGG GTG TGG GAA GCT, probe: ACA CAG GCC AGG TAT TTC AGG TCA GCC; for AR forward primer: GGT GTC ACT ATG GAG CTC TCA CAT, reverse primer: GCA ATC ATT TCT GCT GGC G, probe: CTT CAA AAG AGC CGCTGA AGG GAA ACA G; for PMEPA1 forward primer: CAT GAT CCC CGA GCT GCT, reverse primer: TGA TCT GAA CAA ACT CCA GCT CC, probe: AGG CGG ACA GTC TCC TGC GAA AC; for PCA3 forward primer: CAC ATT TCC AGC CCC TTT AAA TA, reverse primer: GGG CGA GGC TCA TCG AT, probe: GGA AGC ACA GAG ATC CCT GGG AGA AAT G; for ERG forward primer: CAG GTC CTT CTT GCC TCC C, reverse primer: TAT GGA GGC TCC AAT TGA AAC C, probe: TGT CTT TTA TTT CTA GCC CCT TTT GGA ACA GGA; for TMPRSS2-ERG A-type fusion forward primer: CGC GGC AGG AAG CCT TAT, reverse primer: CGC GGT CAT CTC TGT CTT AGC, probe: GCC TAC GGA ACG CCA CAC; for LTF forward primer: AGA GAC TCC CCC ATC CAG TGT AT, reverse primer: CTG TCT TTC GGT CCC GTA GAC TT, probe: CAT TGC GGA AAA CAG; for AMACR: “assay-on-demand” Hs00204885 (Invitrogen, Carlsbad, CA).

    Article Title: Change of voltage-gated potassium channel 1.7 expressions in monocrotaline-induced pulmonary arterial hypertension rat model
    Article Snippet: The mRNA from every sample converted to cDNA (Qiagen, Venlo, The Netherlands). .. Specific PCR primer pair for the studied genes were; 18s ribosomal RNA, forward 5’-GTA ACC CGT TGA ACC TT-3’, reverse 5’-CCA TCC AAT CGG TAG TAG CG-3’; Kv1.7, forward 5’-TCC TTC TAC GGG CTG GGT GC-3’, reverse 5’-TTG AGC CGA GCG AGG AAC GA-3’; Kir6.1, forward 5’-GAG TGA ACT GTC GCA CCA GA-3’; reverse 5’-CGA TCA CCA GAA CTC AGC AA-3’.

    Countercurrent Chromatography:

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. The TaqMan primers and probe for TMPRSS2 were: forward primer: CAC GGA CTG GAT TTA TCG ACA A; reverse primer: CGT CAA GGA CGA AGA CCA TGT, probe: TGA GGG CAG ACG GCT A; for PSA/KLK3 forward primer: CCC ACT GCA TCA GGA ACA AA, reverse primer: GAG CGG GTG TGG GAA GCT, probe: ACA CAG GCC AGG TAT TTC AGG TCA GCC; for AR forward primer: GGT GTC ACT ATG GAG CTC TCA CAT, reverse primer: GCA ATC ATT TCT GCT GGC G, probe: CTT CAA AAG AGC CGCTGA AGG GAA ACA G; for PMEPA1 forward primer: CAT GAT CCC CGA GCT GCT, reverse primer: TGA TCT GAA CAA ACT CCA GCT CC, probe: AGG CGG ACA GTC TCC TGC GAA AC; for PCA3 forward primer: CAC ATT TCC AGC CCC TTT AAA TA, reverse primer: GGG CGA GGC TCA TCG AT, probe: GGA AGC ACA GAG ATC CCT GGG AGA AAT G; for ERG forward primer: CAG GTC CTT CTT GCC TCC C, reverse primer: TAT GGA GGC TCC AAT TGA AAC C, probe: TGT CTT TTA TTT CTA GCC CCT TTT GGA ACA GGA; for TMPRSS2-ERG A-type fusion forward primer: CGC GGC AGG AAG CCT TAT, reverse primer: CGC GGT CAT CTC TGT CTT AGC, probe: GCC TAC GGA ACG CCA CAC; for LTF forward primer: AGA GAC TCC CCC ATC CAG TGT AT, reverse primer: CTG TCT TTC GGT CCC GTA GAC TT, probe: CAT TGC GGA AAA CAG; for AMACR: “assay-on-demand” Hs00204885 (Invitrogen, Carlsbad, CA).

    Concentration Assay:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: Total RNA concentration and purity of samples was measured using the NanoDrop ND‐1000 spectrophotometer (Saveen and Werner AB, Sweden) and RNA integrity was assessed using the Agilent Bioanalyzer 2100 and RNA 6000 Nano Kit (Agilent Technologies, Glostrup, Denmark). .. All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions.

    Generated:

    Article Title: CXCL17 Chemokine-Dependent Mobilization of Memory CXCR8+CD8+ TEM and TRM Cells in the Vaginal Mucosa is Associated with Protection Against Genital Herpes
    Article Snippet: Equal concentrations of RNA were used for the reverse transcription reaction to generate cDNA (Qiagen, Valencia, CA). .. Quantitative real-time PCR (q-PCR) data was generated with a Roche LightCycler® 480 using a Universal Probe Library-based system.

    Article Title: Influence of Aspergillus fumigatus conidia viability on murine pulmonary microRNA and mRNA expression following subchronic inhalation exposure
    Article Snippet: RT2 SYBR Green ROX qPCR Mastermix was used for quantification of cDNA (Qiagen, CA). .. If specific genes included in the response pathways resulted in undetermined expression levels, the genes were removed from the heat maps (generated using R software) that are reported in .

    Polymerase Chain Reaction:

    Article Title: Bis-anthracycline WP760 abrogates melanoma cell growth by transcription inhibition, p53 activation and IGF1R downregulation
    Article Snippet: Paragraph title: PCR array analysis ... Total RNA (3 μg) was reverse-transcribed into cDNA (Qiagen, 330,404) and qPCR was performed using Qiagen reagents (330502) and CFX96 thermocycler (Bio-Rad).

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells
    Article Snippet: .. RNA was isolated and analyzed as indicated above. cDNA was synthesized (Qiagen, RT first strand kit) and the PCR array was performed according to the manufactures protocol (Qiagen, Cat.no. ..

    Article Title: Ovine middle cerebral artery characterization and quantification of ultrastructure and other features: changes with development
    Article Snippet: The mRNA was reverse-transcribed using the Quantitect RT kit, which includes a DNase treatment step before making cDNA (Qiagen). .. Primers were designed using Primer3, a web-based software, and Quantitect SYBR Green PCR Master Mix (Qiagen).

    Article Title: The BH3 mimetic HA14-1 enhances 5-fluorouracil-induced autophagy and type II cell death in oesophageal cancer cells
    Article Snippet: RNA extraction and real-time PCR RNA was extracted, treated with DNase (Ambion, Dublin, Ireland) and cDNA synthesised (Qiagen). .. PCR conditions: 95 °C for 10 min; 50 × for Noxa (β -actin 30 × ) of 95 °C for 10 s, 62 °C for 5 s for Noxa (β -actin 57 °C), 72 °C for 7 s for Noxa (β -actin 8s) (Lightcycler System, Roche, Burgess Hill, West Sussex, UK).

    Article Title: CXCL17 Chemokine-Dependent Mobilization of Memory CXCR8+CD8+ TEM and TRM Cells in the Vaginal Mucosa is Associated with Protection Against Genital Herpes
    Article Snippet: Equal concentrations of RNA were used for the reverse transcription reaction to generate cDNA (Qiagen, Valencia, CA). .. Seventy ng of each cDNA was used per 40 cycle PCR run.

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Paragraph title: PCR array and RT-PCR ... Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA).

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: Real time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was then performed ( ). .. The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA). .. At the end of the run, melting curves were performed for each reaction to ensure the amplification of a single product.

    Article Title: Influence of Aspergillus fumigatus conidia viability on murine pulmonary microRNA and mRNA expression following subchronic inhalation exposure
    Article Snippet: The primers were included in prefabricated PCR plates (RT2 Profiler™ PCR Array Mouse Th1 & Th2 Responses and RT2 Profiler™ PCR Array Mouse Th17 Response). .. RT2 SYBR Green ROX qPCR Mastermix was used for quantification of cDNA (Qiagen, CA).

    Article Title: Change of voltage-gated potassium channel 1.7 expressions in monocrotaline-induced pulmonary arterial hypertension rat model
    Article Snippet: Paragraph title: 6) Polymerase chain reaction analysis ... The mRNA from every sample converted to cDNA (Qiagen, Venlo, The Netherlands).

    Cellular Antioxidant Activity Assay:

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. The TaqMan primers and probe for TMPRSS2 were: forward primer: CAC GGA CTG GAT TTA TCG ACA A; reverse primer: CGT CAA GGA CGA AGA CCA TGT, probe: TGA GGG CAG ACG GCT A; for PSA/KLK3 forward primer: CCC ACT GCA TCA GGA ACA AA, reverse primer: GAG CGG GTG TGG GAA GCT, probe: ACA CAG GCC AGG TAT TTC AGG TCA GCC; for AR forward primer: GGT GTC ACT ATG GAG CTC TCA CAT, reverse primer: GCA ATC ATT TCT GCT GGC G, probe: CTT CAA AAG AGC CGCTGA AGG GAA ACA G; for PMEPA1 forward primer: CAT GAT CCC CGA GCT GCT, reverse primer: TGA TCT GAA CAA ACT CCA GCT CC, probe: AGG CGG ACA GTC TCC TGC GAA AC; for PCA3 forward primer: CAC ATT TCC AGC CCC TTT AAA TA, reverse primer: GGG CGA GGC TCA TCG AT, probe: GGA AGC ACA GAG ATC CCT GGG AGA AAT G; for ERG forward primer: CAG GTC CTT CTT GCC TCC C, reverse primer: TAT GGA GGC TCC AAT TGA AAC C, probe: TGT CTT TTA TTT CTA GCC CCT TTT GGA ACA GGA; for TMPRSS2-ERG A-type fusion forward primer: CGC GGC AGG AAG CCT TAT, reverse primer: CGC GGT CAT CTC TGT CTT AGC, probe: GCC TAC GGA ACG CCA CAC; for LTF forward primer: AGA GAC TCC CCC ATC CAG TGT AT, reverse primer: CTG TCT TTC GGT CCC GTA GAC TT, probe: CAT TGC GGA AAA CAG; for AMACR: “assay-on-demand” Hs00204885 (Invitrogen, Carlsbad, CA).

    Molecular Weight:

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: PCR products were analyzed in a 2% agarose gel, and their sizes were determined by comparison with molecular weight standards after GelRed (Biotium, Hayward, CA, USA) stain; the results were captured with a gel documentation system (DNR Bio-Imaging System, Jerusalem, Israel) using GelCapture Acquisition software (DNR Bio-Imaging Systems). .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    RNA Sequencing Assay:

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells
    Article Snippet: PCR pathway array The human mTOR RT profiler PCR pathway array (SABioscience Qiagen, Cat.no.330231 PAHS-098ZD) was used to validate mTOR regulation identified through RNA seq. .. RNA was isolated and analyzed as indicated above. cDNA was synthesized (Qiagen, RT first strand kit) and the PCR array was performed according to the manufactures protocol (Qiagen, Cat.no.

    Isolation:

    Article Title: Bis-anthracycline WP760 abrogates melanoma cell growth by transcription inhibition, p53 activation and IGF1R downregulation
    Article Snippet: 70% confluency were incubated with 100 nM WP760 or DMSO (control) for 24 h. Subsequently, cells were trypsinized, washed with PBS-, lysed with RLT buffer containing β-ME (Qiagen), and stored at −80 °C until RNA isolation. .. Total RNA (3 μg) was reverse-transcribed into cDNA (Qiagen, 330,404) and qPCR was performed using Qiagen reagents (330502) and CFX96 thermocycler (Bio-Rad).

    Article Title: TGR5 potentiates GLP-1 secretion in response to anionic exchange resins
    Article Snippet: .. Quantitative RT-PCR RNA was isolated from cells using TRIreagent (Ambion), after which cDNA was synthesized (Qiagen). .. Quantitative RT-PCR was performed using SYBR green (Roche) in the Lightcycler 480 II (Roche).

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells
    Article Snippet: .. RNA was isolated and analyzed as indicated above. cDNA was synthesized (Qiagen, RT first strand kit) and the PCR array was performed according to the manufactures protocol (Qiagen, Cat.no. ..

    Article Title: Ovine middle cerebral artery characterization and quantification of ultrastructure and other features: changes with development
    Article Snippet: Isolated mRNA was analyzed using a Nanodrop 2000c (Thermo Scientific, Wilmington, DE) at 260/280-nm wavelength to check for quality and quantity. .. The mRNA was reverse-transcribed using the Quantitect RT kit, which includes a DNase treatment step before making cDNA (Qiagen).

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: .. Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA). .. Predesigned PCR primers for the tested genes were purchased from IDT technologies (Coralville, IA Real-time RT-PCR was performed with 200 ng of total RNA and results were analyzed using the 2−ΔΔCt relative quantification method, after normalization to beta-actin.

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: .. The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. Quantitative gene expression analysis was performed by TaqMan-based QRT-PCR on ABI 7700 (Applied Biosystems, Foster City, CA).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: Total RNA was isolated using TRIzol (Life Technologies) and reverse transcribed into cDNA with oligo dT (Life Technologies). .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    Size-exclusion Chromatography:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: 200 μ L chloroform was added and tubes were shaken vigorously for 15 sec and transferred to 2 mL Eppendorf tubes followed by centrifugation for 15 min at 12000 × g . .. All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions.

    Purification:

    Article Title: CXCL17 is a major chemotactic factor for lung macrophages
    Article Snippet: Total RNA was extracted from mouse tissues using TRIzol (Invitrogen, Carlsbad, CA) and subsequently purified using Qiagen's RNEasy columns and DNase digest (Qiagen). .. Equal concentrations of RNA were used for each tissue sample in a reverse transcription reaction to synthesize cDNA (Qiagen).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: Paragraph title: RNA extraction for RT‐PCR analyses ... All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions.

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Paragraph title: PCR array and RT-PCR ... Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA).

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: Real time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was then performed ( ). .. The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: Paragraph title: Reverse transcriptase-polymerase chain reaction (RT-PCR) and Quantitative PCR (RT-qPCR) ... The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    IA:

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA). .. Predesigned PCR primers for the tested genes were purchased from IDT technologies (Coralville, IA Real-time RT-PCR was performed with 200 ng of total RNA and results were analyzed using the 2−ΔΔCt relative quantification method, after normalization to beta-actin.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells
    Article Snippet: .. RNA was isolated and analyzed as indicated above. cDNA was synthesized (Qiagen, RT first strand kit) and the PCR array was performed according to the manufactures protocol (Qiagen, Cat.no. ..

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. The TaqMan primers and probe for TMPRSS2 were: forward primer: CAC GGA CTG GAT TTA TCG ACA A; reverse primer: CGT CAA GGA CGA AGA CCA TGT, probe: TGA GGG CAG ACG GCT A; for PSA/KLK3 forward primer: CCC ACT GCA TCA GGA ACA AA, reverse primer: GAG CGG GTG TGG GAA GCT, probe: ACA CAG GCC AGG TAT TTC AGG TCA GCC; for AR forward primer: GGT GTC ACT ATG GAG CTC TCA CAT, reverse primer: GCA ATC ATT TCT GCT GGC G, probe: CTT CAA AAG AGC CGCTGA AGG GAA ACA G; for PMEPA1 forward primer: CAT GAT CCC CGA GCT GCT, reverse primer: TGA TCT GAA CAA ACT CCA GCT CC, probe: AGG CGG ACA GTC TCC TGC GAA AC; for PCA3 forward primer: CAC ATT TCC AGC CCC TTT AAA TA, reverse primer: GGG CGA GGC TCA TCG AT, probe: GGA AGC ACA GAG ATC CCT GGG AGA AAT G; for ERG forward primer: CAG GTC CTT CTT GCC TCC C, reverse primer: TAT GGA GGC TCC AAT TGA AAC C, probe: TGT CTT TTA TTT CTA GCC CCT TTT GGA ACA GGA; for TMPRSS2-ERG A-type fusion forward primer: CGC GGC AGG AAG CCT TAT, reverse primer: CGC GGT CAT CTC TGT CTT AGC, probe: GCC TAC GGA ACG CCA CAC; for LTF forward primer: AGA GAC TCC CCC ATC CAG TGT AT, reverse primer: CTG TCT TTC GGT CCC GTA GAC TT, probe: CAT TGC GGA AAA CAG; for AMACR: “assay-on-demand” Hs00204885 (Invitrogen, Carlsbad, CA).

    RNA Extraction:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: Paragraph title: RNA extraction for RT‐PCR analyses ... All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions.

    Article Title: The BH3 mimetic HA14-1 enhances 5-fluorouracil-induced autophagy and type II cell death in oesophageal cancer cells
    Article Snippet: .. RNA extraction and real-time PCR RNA was extracted, treated with DNase (Ambion, Dublin, Ireland) and cDNA synthesised (Qiagen). .. Each real-time PCR reaction contained 0.5 μ of each primer, MgCl2 (Noxa 4 m, β -actin 3 m), 1 × LightCycler FastStart DNA Master SYBR Green and template cDNA.

    Software:

    Article Title: Ovine middle cerebral artery characterization and quantification of ultrastructure and other features: changes with development
    Article Snippet: The mRNA was reverse-transcribed using the Quantitect RT kit, which includes a DNase treatment step before making cDNA (Qiagen). .. Primers were designed using Primer3, a web-based software, and Quantitect SYBR Green PCR Master Mix (Qiagen).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: PCR products were analyzed in a 2% agarose gel, and their sizes were determined by comparison with molecular weight standards after GelRed (Biotium, Hayward, CA, USA) stain; the results were captured with a gel documentation system (DNR Bio-Imaging System, Jerusalem, Israel) using GelCapture Acquisition software (DNR Bio-Imaging Systems). .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    Article Title: Influence of Aspergillus fumigatus conidia viability on murine pulmonary microRNA and mRNA expression following subchronic inhalation exposure
    Article Snippet: RT2 SYBR Green ROX qPCR Mastermix was used for quantification of cDNA (Qiagen, CA). .. If specific genes included in the response pathways resulted in undetermined expression levels, the genes were removed from the heat maps (generated using R software) that are reported in .

    Real-time Polymerase Chain Reaction:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions. .. Finally, 91 μ L RNase‐free water was mixed with each cDNA reaction. cDNA for Fluidigm qPCR (see below) analysis was prepared by reverse transcription of 500 ng duplicate samples of extracted total RNA using the QuantiTECT Reverse Transcription kit (Qiagen) as described previously (Skovgaard et al. ).

    Article Title: Bis-anthracycline WP760 abrogates melanoma cell growth by transcription inhibition, p53 activation and IGF1R downregulation
    Article Snippet: .. Total RNA (3 μg) was reverse-transcribed into cDNA (Qiagen, 330,404) and qPCR was performed using Qiagen reagents (330502) and CFX96 thermocycler (Bio-Rad). .. RNA isolation was performed using RNeasy Mini Kit (Qiagen) following the manufacturer’s protocol.

    Article Title: Ovine middle cerebral artery characterization and quantification of ultrastructure and other features: changes with development
    Article Snippet: Paragraph title: Real-time PCR. ... The mRNA was reverse-transcribed using the Quantitect RT kit, which includes a DNase treatment step before making cDNA (Qiagen).

    Article Title: The BH3 mimetic HA14-1 enhances 5-fluorouracil-induced autophagy and type II cell death in oesophageal cancer cells
    Article Snippet: .. RNA extraction and real-time PCR RNA was extracted, treated with DNase (Ambion, Dublin, Ireland) and cDNA synthesised (Qiagen). .. Each real-time PCR reaction contained 0.5 μ of each primer, MgCl2 (Noxa 4 m, β -actin 3 m), 1 × LightCycler FastStart DNA Master SYBR Green and template cDNA.

    Article Title: CXCL17 Chemokine-Dependent Mobilization of Memory CXCR8+CD8+ TEM and TRM Cells in the Vaginal Mucosa is Associated with Protection Against Genital Herpes
    Article Snippet: Equal concentrations of RNA were used for the reverse transcription reaction to generate cDNA (Qiagen, Valencia, CA). .. Quantitative real-time PCR (q-PCR) data was generated with a Roche LightCycler® 480 using a Universal Probe Library-based system.

    Article Title: Silencing growth hormone receptor inhibits estrogen receptor negative breast cancer through ATP-binding cassette sub-family G member 2
    Article Snippet: Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA) and reverse transcribed to cDNA (Qiagen, Valencia, CA). .. Briefly, total RNA was isolated from cells and cDNA was prepared using RT2 First Strand kit (SA Biosciences, Frederick, MD) and SYBR green based real-time PCR was carried out using StepOnePlus Real-Time PCR machine (Applied Biosystems, Foster City, CA).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA). .. At the end of the run, melting curves were performed for each reaction to ensure the amplification of a single product.

    Article Title: Influence of Aspergillus fumigatus conidia viability on murine pulmonary microRNA and mRNA expression following subchronic inhalation exposure
    Article Snippet: .. RT2 SYBR Green ROX qPCR Mastermix was used for quantification of cDNA (Qiagen, CA). .. Data were normalized by gene expression relative to the levels of glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) for the Th1–Th2 response pathway and beta-glucuronidase ( Gusb ) for the Th17 response pathway.

    Article Title: Intrauterine inflammation alters cardiopulmonary and cerebral haemodynamics at birth in preterm lambs
    Article Snippet: Total RNA was extracted from lung tissue and reverse transcribed to produce cDNA (Qiagen, Melbourne, Victoria, Australia). .. The mRNA levels for CTGF, CYR-61 EGR-1 and IL-1β, IL-8 and IL-6 were determined using quantitative real-time PCR as described previously ( ).

    Negative Control:

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: As negative control for PCR amplification, reactions with RNA in the absence of reverse transcriptase as well human peripheral blood and mouse embryonic fibroblast cells were included. .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    Agarose Gel Electrophoresis:

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: PCR products were analyzed in a 2% agarose gel, and their sizes were determined by comparison with molecular weight standards after GelRed (Biotium, Hayward, CA, USA) stain; the results were captured with a gel documentation system (DNR Bio-Imaging System, Jerusalem, Israel) using GelCapture Acquisition software (DNR Bio-Imaging Systems). .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

    Homogenization:

    Article Title: Bis-anthracycline WP760 abrogates melanoma cell growth by transcription inhibition, p53 activation and IGF1R downregulation
    Article Snippet: Total RNA (3 μg) was reverse-transcribed into cDNA (Qiagen, 330,404) and qPCR was performed using Qiagen reagents (330502) and CFX96 thermocycler (Bio-Rad). .. An additional homogenization step (QIAshredder) or homogenization by passing through a 21-gauge needle, and DNase I (Qiagen) digestion were also performed.

    Laser Capture Microdissection:

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: Total RNA was isolated from the LCM specimens using the MicroRNA kit (Stratagene, La Jolla, CA). .. The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA).

    Spectrophotometry:

    Article Title: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets
    Article Snippet: Total RNA concentration and purity of samples was measured using the NanoDrop ND‐1000 spectrophotometer (Saveen and Werner AB, Sweden) and RNA integrity was assessed using the Agilent Bioanalyzer 2100 and RNA 6000 Nano Kit (Agilent Technologies, Glostrup, Denmark). .. All RNA integrity values (RIN) were between 5.9 and 8.5. cDNA (Qiagen array, see below) was synthesized using the RT2 First Strand Kit provided by Qiagen, using the Stratagene MX3000p according to the manufacturer's instructions.

    Activation Assay:

    Article Title: Differential Efficacy of 2 Vibrating Orthodontic Devices to Alter the Cellular Response in Osteoblasts, Fibroblasts, and Osteoclasts
    Article Snippet: RNA was reverse transcribed to complementary DNA (Qiagen). .. For osteoclasts, we quantified phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), receptor activator of nuclear factor κ B (RANK), and nuclear factor of activated T cells 1 (NFATC1) with PI3K as indicator of the Akt signaling pathway (reflecting osteoclast activation/proliferation) and NFATC1 and RANK as indicators of osteoclast differentiation during osteoclastogenesis.

    CTG Assay:

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA). .. The TaqMan primers and probe for TMPRSS2 were: forward primer: CAC GGA CTG GAT TTA TCG ACA A; reverse primer: CGT CAA GGA CGA AGA CCA TGT, probe: TGA GGG CAG ACG GCT A; for PSA/KLK3 forward primer: CCC ACT GCA TCA GGA ACA AA, reverse primer: GAG CGG GTG TGG GAA GCT, probe: ACA CAG GCC AGG TAT TTC AGG TCA GCC; for AR forward primer: GGT GTC ACT ATG GAG CTC TCA CAT, reverse primer: GCA ATC ATT TCT GCT GGC G, probe: CTT CAA AAG AGC CGCTGA AGG GAA ACA G; for PMEPA1 forward primer: CAT GAT CCC CGA GCT GCT, reverse primer: TGA TCT GAA CAA ACT CCA GCT CC, probe: AGG CGG ACA GTC TCC TGC GAA AC; for PCA3 forward primer: CAC ATT TCC AGC CCC TTT AAA TA, reverse primer: GGG CGA GGC TCA TCG AT, probe: GGA AGC ACA GAG ATC CCT GGG AGA AAT G; for ERG forward primer: CAG GTC CTT CTT GCC TCC C, reverse primer: TAT GGA GGC TCC AAT TGA AAC C, probe: TGT CTT TTA TTT CTA GCC CCT TTT GGA ACA GGA; for TMPRSS2-ERG A-type fusion forward primer: CGC GGC AGG AAG CCT TAT, reverse primer: CGC GGT CAT CTC TGT CTT AGC, probe: GCC TAC GGA ACG CCA CAC; for LTF forward primer: AGA GAC TCC CCC ATC CAG TGT AT, reverse primer: CTG TCT TTC GGT CCC GTA GAC TT, probe: CAT TGC GGA AAA CAG; for AMACR: “assay-on-demand” Hs00204885 (Invitrogen, Carlsbad, CA).

    Article Title: Change of voltage-gated potassium channel 1.7 expressions in monocrotaline-induced pulmonary arterial hypertension rat model
    Article Snippet: The mRNA from every sample converted to cDNA (Qiagen, Venlo, The Netherlands). .. Specific PCR primer pair for the studied genes were; 18s ribosomal RNA, forward 5’-GTA ACC CGT TGA ACC TT-3’, reverse 5’-CCA TCC AAT CGG TAG TAG CG-3’; Kv1.7, forward 5’-TCC TTC TAC GGG CTG GGT GC-3’, reverse 5’-TTG AGC CGA GCG AGG AAC GA-3’; Kir6.1, forward 5’-GAG TGA ACT GTC GCA CCA GA-3’; reverse 5’-CGA TCA CCA GAA CTC AGC AA-3’.

    Staining:

    Article Title: Quantitative Expression of TMPRSS2 Transcript in Prostate Tumor Cells Reflects TMPRSS2-ERG Fusion Status
    Article Snippet: Benign and malignant cells were laser capture microdissected (LCM) by a pathologist from optimum cutting temperature (OCT) medium-embedded and H & E stained frozen prostate sections of RP specimens (2000 laser shots per sample). .. The isolated total RNA was converted to cDNA (Sensiscript, Qiagen, Valencia, CA).

    Article Title: Markers of Pluripotency in Human Amniotic Epithelial Cells and Their Differentiation to Progenitor of Cortical Neurons
    Article Snippet: PCR products were analyzed in a 2% agarose gel, and their sizes were determined by comparison with molecular weight standards after GelRed (Biotium, Hayward, CA, USA) stain; the results were captured with a gel documentation system (DNR Bio-Imaging System, Jerusalem, Israel) using GelCapture Acquisition software (DNR Bio-Imaging Systems). .. The qPCR-amplification parameters were the same as for end-point PCR reactions, but using KAPASYBR® FAST Universal qPCR Master Mix (KAPA Biosystems, Boston, MA, USA). qPCR was carried out in a Rotor-Gene Q 7000 thermocycler using 800 ng of cDNA (Qiagen, Valencia, CA, USA).

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    For real time quantitative PCR analysis of control gene fusion gene or mutated gene transcripts Kit contents For 8 reactions 1 tube per standard dilution Benefits Sensitive quantification EAC standardized
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    Qiagen cdna synthesis total rna
    Example of reticulocyte <t>cDNA</t> quality control. Agarose gel electrophoresis of RT ‐ PCR products (image inverted, black/white) obtained using reticulocyte cDNA from patients ( HA patient, N62, and HS patient, C14) and controls (healthy individuals: C1 and C2) and primers encoding sequences of the following genes: CD 45 ( PTPRC gene; 217 bp)—leukocyte marker; β‐globin ( HBB gene; 397 bp)—erythroid marker; integrin‐β3 ( ITGB 3 gene; 198 bp)—platelet marker. As the “low abundance” gene transcript PPARA (peroxisome proliferator activated receptor alpha; 324 bp) was chosen and was taken from the end of the reticulocyte cDNA library ( NCBI , Library 11923). For the loading control β‐actin primers ( ACTB gene; 479 bp) were used. All primer sequences used in this experiment are included in Supporting Information Table S2 . As a standard “GeneRuler 100 bp DNA Ladder” (Thermo Fisher Scientific, Waltham, MA, USA) was used. Results show no leukocyte or thrombocyte <t>RNA</t> contamination for all analysed cases. The lowest band corresponds to primers
    Cdna Synthesis Total Rna, supplied by Qiagen, used in various techniques. Bioz Stars score: 94/100, based on 266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen cdna construction
    RT–PCR amplification products from different tissues using the primer set CCV-mut-F/-R. Lane 1: Epstein Barr virus–transformed B-lymphocytes cell <t>cDNA</t> preparations showing the 218 bp transcripts. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and the acceptor splice site: TTT CCT CCA G^G CTG CAC CA-3′. Lane 2: A 500 bp genomic product. Lane 3: Universal Human reference <t>RNA</t> (mixed carcinoma) shows the 324 bp product. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and acceptor splice site: CTC TCC CCA G^C CAG CGC CCT-3′. Lane 4: cDNA from a human fetal eye from DNase-treated RNA. A 324 bp product (arrow) is seen. Lane 5: cDNA from human lens epithelial cells (HLEpiC cat. No. 6554 lot no. 2531, ScienCell) not DNase treated. A genomic DNA (500 bp) and a 324 bp PCR product (arrow) are seen from the spliced RP1–140A9.1 . Lane 6: 100 bp ladder.
    Cdna Construction, supplied by Qiagen, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen cdna generation
    Overexpression of Tmie causes increased bundle expression of Tmc1-GFP and Tmc2b-GFP. (A) Semi-quantitative <t>PCR</t> of tmie <t>cDNA</t> using primers within exon 4 to detect transcripts from both transgene and the endogenous gene. We generated cDNA from RNA extracted from 5 dpf whole larvae. Larvae carrying Tg(myo6b : tmie-p2A-NLS(mCherry)) had 6.5-fold more tmie transcript than wild type non-transgenic siblings. Products are from 40 cycles. Less PCR reaction (2.5x less) was loaded for gapdh to avoid saturation. (B) Confocal images of the lateral cristae in 4 dpf tmie ru1000 larvae co-expressing two transgenes, tmc1-GFP (upper panel) and tmie-p2A-NLS(mCherry) (lower panel). The p2A linker is a self-cleaving peptide that results in equimolar translation of Tmie and nuclear mCherry. (C) Plot of the integrated density of Tmc1-GFP fluorescence/crista in the ROI, expressed in arbitrary units; significance determined by one-way ANOVA. (D-E) Same as B-C except using tmc2b-GFP instead of tmc1-GFP . All statistics are mean ± SD. Scale bars are 5μm.
    Cdna Generation, supplied by Qiagen, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Example of reticulocyte cDNA quality control. Agarose gel electrophoresis of RT ‐ PCR products (image inverted, black/white) obtained using reticulocyte cDNA from patients ( HA patient, N62, and HS patient, C14) and controls (healthy individuals: C1 and C2) and primers encoding sequences of the following genes: CD 45 ( PTPRC gene; 217 bp)—leukocyte marker; β‐globin ( HBB gene; 397 bp)—erythroid marker; integrin‐β3 ( ITGB 3 gene; 198 bp)—platelet marker. As the “low abundance” gene transcript PPARA (peroxisome proliferator activated receptor alpha; 324 bp) was chosen and was taken from the end of the reticulocyte cDNA library ( NCBI , Library 11923). For the loading control β‐actin primers ( ACTB gene; 479 bp) were used. All primer sequences used in this experiment are included in Supporting Information Table S2 . As a standard “GeneRuler 100 bp DNA Ladder” (Thermo Fisher Scientific, Waltham, MA, USA) was used. Results show no leukocyte or thrombocyte RNA contamination for all analysed cases. The lowest band corresponds to primers

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Efficient method for isolation of reticulocyte RNA from healthy individuals and hemolytic anaemia patients, et al. Efficient method for isolation of reticulocyte RNA from healthy individuals and hemolytic anaemia patients

    doi: 10.1111/jcmm.13951

    Figure Lengend Snippet: Example of reticulocyte cDNA quality control. Agarose gel electrophoresis of RT ‐ PCR products (image inverted, black/white) obtained using reticulocyte cDNA from patients ( HA patient, N62, and HS patient, C14) and controls (healthy individuals: C1 and C2) and primers encoding sequences of the following genes: CD 45 ( PTPRC gene; 217 bp)—leukocyte marker; β‐globin ( HBB gene; 397 bp)—erythroid marker; integrin‐β3 ( ITGB 3 gene; 198 bp)—platelet marker. As the “low abundance” gene transcript PPARA (peroxisome proliferator activated receptor alpha; 324 bp) was chosen and was taken from the end of the reticulocyte cDNA library ( NCBI , Library 11923). For the loading control β‐actin primers ( ACTB gene; 479 bp) were used. All primer sequences used in this experiment are included in Supporting Information Table S2 . As a standard “GeneRuler 100 bp DNA Ladder” (Thermo Fisher Scientific, Waltham, MA, USA) was used. Results show no leukocyte or thrombocyte RNA contamination for all analysed cases. The lowest band corresponds to primers

    Article Snippet: 2.4 RNA isolation and cDNA synthesis Total RNA was extracted from the reticulocyte‐rich suspension and collected using the miRNeasy Mini Kit (Qiagen, Hilden, Germany), according to the manufacturer's recommendation.

    Techniques: Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Marker, cDNA Library Assay

    RT–PCR amplification products from different tissues using the primer set CCV-mut-F/-R. Lane 1: Epstein Barr virus–transformed B-lymphocytes cell cDNA preparations showing the 218 bp transcripts. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and the acceptor splice site: TTT CCT CCA G^G CTG CAC CA-3′. Lane 2: A 500 bp genomic product. Lane 3: Universal Human reference RNA (mixed carcinoma) shows the 324 bp product. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and acceptor splice site: CTC TCC CCA G^C CAG CGC CCT-3′. Lane 4: cDNA from a human fetal eye from DNase-treated RNA. A 324 bp product (arrow) is seen. Lane 5: cDNA from human lens epithelial cells (HLEpiC cat. No. 6554 lot no. 2531, ScienCell) not DNase treated. A genomic DNA (500 bp) and a 324 bp PCR product (arrow) are seen from the spliced RP1–140A9.1 . Lane 6: 100 bp ladder.

    Journal: Molecular Vision

    Article Title: A splice-site variant in the lncRNA gene RP1-140A9.1 cosegregates in the large Volkmann cataract family

    doi:

    Figure Lengend Snippet: RT–PCR amplification products from different tissues using the primer set CCV-mut-F/-R. Lane 1: Epstein Barr virus–transformed B-lymphocytes cell cDNA preparations showing the 218 bp transcripts. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and the acceptor splice site: TTT CCT CCA G^G CTG CAC CA-3′. Lane 2: A 500 bp genomic product. Lane 3: Universal Human reference RNA (mixed carcinoma) shows the 324 bp product. Donor splice site: 5′-AGA CCT CCA G^G TGA GGA AGG and acceptor splice site: CTC TCC CCA G^C CAG CGC CCT-3′. Lane 4: cDNA from a human fetal eye from DNase-treated RNA. A 324 bp product (arrow) is seen. Lane 5: cDNA from human lens epithelial cells (HLEpiC cat. No. 6554 lot no. 2531, ScienCell) not DNase treated. A genomic DNA (500 bp) and a 324 bp PCR product (arrow) are seen from the spliced RP1–140A9.1 . Lane 6: 100 bp ladder.

    Article Snippet: RNA isolation, cDNA construction, and qPCR Total RNA was extracted from Epstein Barr transformed B-lymphocytes [ ], whole venous blood was taken from five healthy and affected individuals individuals (later frozen) and from human eye [ ] using an RNeasy mini kit from Qiagen (Cat No./ID: 74,104; Hilden, Germany).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Transformation Assay, CTG Assay, Polymerase Chain Reaction

    Overexpression of Tmie causes increased bundle expression of Tmc1-GFP and Tmc2b-GFP. (A) Semi-quantitative PCR of tmie cDNA using primers within exon 4 to detect transcripts from both transgene and the endogenous gene. We generated cDNA from RNA extracted from 5 dpf whole larvae. Larvae carrying Tg(myo6b : tmie-p2A-NLS(mCherry)) had 6.5-fold more tmie transcript than wild type non-transgenic siblings. Products are from 40 cycles. Less PCR reaction (2.5x less) was loaded for gapdh to avoid saturation. (B) Confocal images of the lateral cristae in 4 dpf tmie ru1000 larvae co-expressing two transgenes, tmc1-GFP (upper panel) and tmie-p2A-NLS(mCherry) (lower panel). The p2A linker is a self-cleaving peptide that results in equimolar translation of Tmie and nuclear mCherry. (C) Plot of the integrated density of Tmc1-GFP fluorescence/crista in the ROI, expressed in arbitrary units; significance determined by one-way ANOVA. (D-E) Same as B-C except using tmc2b-GFP instead of tmc1-GFP . All statistics are mean ± SD. Scale bars are 5μm.

    Journal: PLoS Genetics

    Article Title: Subunits of the mechano-electrical transduction channel, Tmc1/2b, require Tmie to localize in zebrafish sensory hair cells

    doi: 10.1371/journal.pgen.1007635

    Figure Lengend Snippet: Overexpression of Tmie causes increased bundle expression of Tmc1-GFP and Tmc2b-GFP. (A) Semi-quantitative PCR of tmie cDNA using primers within exon 4 to detect transcripts from both transgene and the endogenous gene. We generated cDNA from RNA extracted from 5 dpf whole larvae. Larvae carrying Tg(myo6b : tmie-p2A-NLS(mCherry)) had 6.5-fold more tmie transcript than wild type non-transgenic siblings. Products are from 40 cycles. Less PCR reaction (2.5x less) was loaded for gapdh to avoid saturation. (B) Confocal images of the lateral cristae in 4 dpf tmie ru1000 larvae co-expressing two transgenes, tmc1-GFP (upper panel) and tmie-p2A-NLS(mCherry) (lower panel). The p2A linker is a self-cleaving peptide that results in equimolar translation of Tmie and nuclear mCherry. (C) Plot of the integrated density of Tmc1-GFP fluorescence/crista in the ROI, expressed in arbitrary units; significance determined by one-way ANOVA. (D-E) Same as B-C except using tmc2b-GFP instead of tmc1-GFP . All statistics are mean ± SD. Scale bars are 5μm.

    Article Snippet: cDNA generation by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and semi-quantitative PCR For and , we extracted total RNA using the RNeasy mini kit (Qiagen).

    Techniques: Over Expression, Expressing, Real-time Polymerase Chain Reaction, Generated, Transgenic Assay, Polymerase Chain Reaction, Fluorescence