Rnazol rt

Manufactured by Euromedex

RNAzol-RT is a reagent for the isolation and purification of total RNA from a variety of biological samples. It is a monophasic solution that combines phenol and guanidinium thiocyanate to facilitate the lysis of cells and the subsequent separation of RNA from DNA and proteins.

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Lab products found in correlation

2 protocols using rnazol rt

Comprehensive miRNA Profiling and Regulation

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Cell culture medium and reagents, oligofectamine, lipofectamine 2000, random hexamers, turbo DNA-free kit, miRNA assay kit, miRNA reverse transcription kit, synthetic mature and pre-mature miRNAs, anti-miR-200 oligonucleotides, MCPiP1 siRNA, PCR TaqMan probes, Go TaqH hot start DNA polymerase and Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV) were provided by Life Technologies. Gemcitabine (GEM), 5-Azacytidine (5-AZA) and anti-β-actine antibody were from Sigma-Aldrich. Enhanced chemiluminescent substrate was from Millipore. Protease inhibitor cocktail was provided by Roche Life Science. Kapa Fast Sybr qPCR Mix was from Clinisciences. RNAzol-RT, RanQ69L-GTP and anti-Dicer1 antibody were from Euromedex. GoTaq G2 DNA polymerase, Go Script reverse transcriptase and RNAsin RNase inhibitor were from Promega. PCR, qPCR and RT specific primers were obtained from Eurogentec. PolyA polymerase tailing kit, anti-MCPiP1, anti-Histone H1 and anti-calnexin antibodies were from Tebu-Bio. Anti-XPO₅ antibody was from Cell Signalling. Protein A/G plus agarose was provided by Santa Cruz. pDestmycDICER1 expression vector was a gift from Thomas Tuschi (Addgene plasmid # 19873).

Boudouresque F., Siret C., Dobric A., Silvy F., Soubeyran P., Iovanna J., Lombardo D, & Berthois Y. (2018). Ribonuclease MCPiP1 contributes to the loss of micro-RNA-200 family members in pancreatic cancer cells. Oncotarget, 9(89), 35941-35961.

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Quantifying A3G mRNA Expression

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Twenty-four hours post-transfection, total RNA was isolated from HEK 293T cells using RNAzol®RT (Euromedex). After RNase-free DNase treatment (TURBO DNA-free kit, Invitrogen), total RNA (1 µg) was reverse-transcribed using the iScript TM Reverse Transcription Supermix (Bio-Rad) as recommended by the manufacturer. Subsequent qPCR analysis was performed using the Maxima TM SYBR Green qPCR Master Mix (ThermoFisher) and was monitored on a CFX Real Time System (Bio-Rad). Gene-specific primers for A3G and β -actin are detailed in Table 1. The A3G mRNA levels were normalized to those of actin mRNA and relative quantification was determined using the standard curve-based method.

Libre C., Seissler T., Guerrero S., Batisse J., Verriez C., Stupfler B., Gilmer O., Cabrera-Rodríguez R., Cimarelli A., Valenzuela-Fernández A., Marquet R., & Etienne L. (2021). A conserved uORF regulates APOBEC3G translation and is targeted by HIV-1 Vif protein to repress the antiviral factor.

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