Identifikasjon av Pålitelige Referanse Gener i den Menneskelige Tarmparasitten Blastocystis ST7-B ved Bruk av Semi-Kvantitativ Revers Rranskriptase PRC: En Kronisk Vurdering

Abstract

The aim of this study was to Evaluate the stability of RNA expression of the candidate genes during the growth of Blastocystis ST7-B. For this we also evaluated the suitability of 12 reference genes as primers for Blastocystis. The primary method utilized in the study was semi-quantitative reverse transcriptase PCR, sqRT-PCR. After a process of elimination, evaluation the ability for each primer pair to amplify genomic DNA and complementary DNA of ST7-B, the final reference genes chosen to be used as primers were Actin, Tubulin, Ribosomal Protein L10 Family, TATA-BOX Binding Protein, Retrieval of Early ER Protein 1, Hypoxanthine Phosphoribosyltransferase, and Glyceraldehyde-3-phosphate dehydrogenase. RER1 and RPLP0-1 were concluded to be the most stable, and recommended to be used in future studies regarding the RNA expression in Blastocystis ST7-B. While GAPDH was proven to be the least stable. Blastocystis – housekeeping genes – reference genes – transcriptomics – RNA expression

The aim of this study was to Evaluate the stability of RNA expression of the candidate genes during the growth of Blastocystis ST7-B. For this we also evaluated the suitability of 12 reference genes as primers for Blastocystis. The primary method utilized in the study was semi-quantitative reverse transcriptase PCR, sqRT-PCR. After a process of elimination, evaluation the ability for each primer pair to amplify genomic DNA and complementary DNA of ST7-B, the final reference genes chosen to be used as primers were Actin, Tubulin, Ribosomal Protein L10 Family, TATA-BOX Binding Protein, Retrieval of Early ER Protein 1, Hypoxanthine Phosphoribosyltransferase, and Glyceraldehyde-3-phosphate dehydrogenase. RER1 and RPLP0-1 were concluded to be the most stable, and recommended to be used in future studies regarding the RNA expression in Blastocystis ST7-B. While GAPDH was proven to be the least stable. Blastocystis – housekeeping genes – reference genes – transcriptomics – RNA expression