Oligo dt 16 18

Manufactured by Thermo Fisher Scientific

Oligo (dT)16–18 is a synthetic oligonucleotide composed of 16 to 18 deoxythymidine nucleotides. It is primarily used as a primer for reverse transcription of polyadenylated RNA.

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Lab products found in correlation

Superscript 2, by Thermo Fisher Scientific (4 mentions) Rnase h, by Thermo Fisher Scientific (4 mentions) Dntps, by Thermo Fisher Scientific (4 mentions) Rnasin, by Promega (4 mentions) Dithiothreitol dtt, by Thermo Fisher Scientific (2 mentions) Dithiothreitol (dtt), by Thermo Fisher Scientific (2 mentions) Rna stat 60, by Tel-Test (2 mentions) Quantitect sybr green pcr reagent, by Qiagen (1 mentions)

Spelling variants of this product

Oligo dT (457 citations) Oligo(dT)18 (51 citations)

4 protocols using oligo dt 16 18

RNA Extraction and cDNA Synthesis

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Cells or tissues were homogenized in RNA Stat60^® and total RNA extracted using standard phenol-chloroform protocols followed by DNase treatment of the extracted RNA using RNA-II purification kit (Nachery-Nagel). A total of 100 ng of RNA per sample was converted into cDNA using Superscript II (Life Technologies) at 42 °C for 50 min, 70 °C 15 min, in the presence of 5 uM oligo (dT)_16–18, 5 mM Dithiothreitol (DTT), 0.5 mM dNTPs (all Life Technologies), 8 U RNAsin (Promega), 50 mM Tris-HCl pH 8.3, 75 mM KCl and 3 mM MgCl₂. The cDNA was treated with 2.5 U RNase H (Affymetrix) at 37 °C for 20 min to remove any remaining RNA residues.

Mimche P.N., Lee C.M., Mimche S.M., Thapa M., Grakoui A., Henkemeyer M, & Lamb T.J. (2018). EphB2 receptor tyrosine kinase promotes hepatic fibrogenesis in mice via activation of hepatic stellate cells. Scientific Reports, 8, 2532.

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RNA Extraction and cDNA Synthesis

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Cells or tissue were homogenized in RNA-Stat60 (Tel-Test Inc) and total RNA was extracted using standard phenol-chloroform protocols followed by DNAse treatment of the RNA extracted using RNA-II purification kit (Nachery-Nagel). A total of 100ng of RNA per sample was converted into cDNA using Superscript II (Life Technologies) at 42°C for 50 min., 70°C for 15 min., in the presence of 5uM oligo (dT)_16-18, 5mM Dithiothreitol (DTT), 0.5mM dNTPs (all from Life Technologies), 8U RNAsin (Promega), 50mM Tris-HCl pH8.3, 75mM KCl and 3mM MgCl₂. The cDNA was treated with 2.5U RNAse H (Affymetrix) at 37°C for 20min to remove any remaining RNA residues.

Darling T.K., Mimche P.N., Bray C., Umaru B., Brady L.M., Stone C., Eboumbou Moukoko C.E., Lane T.E., Ayong L.S, & Lamb T.J. (2020). EphA2 contributes to disruption of the blood-brain barrier in cerebral malaria. PLoS Pathogens, 16(1), e1008261.

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RNA Extraction and cDNA Synthesis

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Cells or tissue were homogenized in RNA Stat60® and total RNA extracted using standard phenol-chloroform protocols followed by DNase treatment of RNA extracted using RNA-II purification kit (Nachery-Nagel). A total of 100ng of RNA per sample was converted into cDNA using Superscript II (Life Technologies) at 42°C for 50min, 70°C 15min, in the presence of 5uM oligo (dT)_16–18, 5mM Dithiothreitol (DTT), 0.5mM dNTPs (all Life Technologies), 8U RNAsin (Promega), 50mM Tris-HCl pH8.3, 75mM KCl and 3mM MgCl₂. The cDNA was treated with 2.5U RNAse H (Affymetrix) at 37°C for 20min to remove any remaining RNA residues.

Mimche P.N., Brady L.M., Bray C.F., Mimche S.M., Thapa M., King T.P., Quicke K., McDermott C.D., Lee C.M., Grakoui A., Morgan E.T, & Lamb T.J. (2015). The receptor tyrosine kinase EphB2 promotes hepatic fibrosis in mice. Hepatology (Baltimore, Md.), 62(3), 900-914.

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RNA Extraction and qPCR Analysis Protocol

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Cells were homogenized in RNA Stat60^® (Tel-Test Inc., TX, USA) and total RNA extracted using standard phenol-chloroform protocols followed by DNase treatment of RNA extracted using Nucleospin RNA-II purification kit (Nachery-Nagel). A total of 100ng of RNA per sample was converted into cDNA using Superscript II (Life Technologies) at 42°C for 50min, 70°C 15min, in the presence of 5μM oligo (dT)_16-18, 5mM Dithiothreitol (DTT), 0.5mM dNTPs (all Life Technologies), 8U RNAsin (Promega), 50mM Tris-HCl pH8.3, 75mM KCl and 3mM MgCl₂. The cDNA was treated with 2.5U RNAse H (Affymetrix) at 37°C for 20min to remove any remaining RNA residues. Real-time qPCR reactions were performed using Quantitect SYBR Green PCR reagent (Qiagen). PCR amplification was performed with 5μl cDNA sample (diluted 1:10), 2μM of each primer and 7μl of QPCR SYBR green mix. Plates were run using an Applied BioSystems FAST 7000 Sequence detection system (ABI Prism FAST 7000). Primer sequences are shown in supporting information S1 Table. Transcripts were normalized to two different housekeeping genes (Ubiquitin and β-actin) and expression levels calculated using the 2^-ΔΔCt method [25 (link)].

Mimche P.N., Brady L.M., Keeton S., Fenne D.S., King T.P., Quicke K.M., Hudson L.E, & Lamb T.J. (2015). Expression of the Receptor Tyrosine Kinase EphB2 on Dendritic Cells Is Modulated by Toll-Like Receptor Ligation but Is Not Required for T Cell Activation. PLoS ONE, 10(9), e0138835.

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