Sukohar, Asep and HERAWATI, HENING and WITARTO, ARIEF B and T SIBERO, HENDRA and SETIAWAN, SETIAWAN and F, FIRMAN and WIRAKUSUMA, WIRAKUSUMA and SASTRAMIHARDJA S., HERRY (2014) MIR-423-3P USED AS REFERENCE GENE FOR MIRNA 146 A IN CELL LINES HEP-G2. International Journal of Pharmacy and Pharmaceutical Sciences, 6 (8). pp. 554-557. ISSN 0975-1491

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do a series of tests in order to get a reference genes. Based on the literature there are 7 candidates recommended reference genes; mir-423-3p, 423-5p, 191, 103, 21, 16, and let-7a. We conducted using four candidates reference gene; mir-423-3p, 103, 16, and 21. Methods: In vitro study was performed in Hep-G2 cells. The samples were divided into control group and experiment group treated with 727μM chlorogenic acid. Samples were analyzed at 0, 2, 8, 18, and 24 hours after being treated with chlorogenic acid. Total RNA was isolated from Hep-G2 with RNA extraction kit (miRCURYTM RNA Isolation Kit-Cell and plant Exiqon, Code Number 300110) and reverse transcribed to cDNA with Primerscript RT Reagent Kit (miRCURY LNATM Universal RT microRNA PCR, Polyadenylation and cDNA synthesis kit Exiqon, Code Number 203300). The primers for miRNA 146 A were Code Number 204688 from Exiqon (forward and reverse), and quantitative real time reverse transcription polymerase chain reaction was performed using SYBR Master Mix with Code Number 203450 (Exicon). MiRNA profiling was performed in four pairs of mir, consist of; mir-423-3p, 103, 21, and mir-16. By using the mean expression value of all expressed mir, we identified the most stable candidate reference genes for subsequent validation with normfinder software.

Item Type: Article
Subjects: R Medicine > RM Therapeutics. Pharmacology
Divisions: Fakultas Kedokteran (FK) > Prodi Pendidikan Dokter
Depositing User: Dr. ASEP SUKOHAR
Date Deposited: 18 Jul 2016 07:49
Last Modified: 31 Aug 2016 02:30

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