Genes, Vol. 10, Pages 376: Identification and Validation of Novel Reference Genes in Acute Lymphoblastic Leukemia for Droplet Digital PCR
Genes, Vol. 10, Pages 376: Identification and Validation of Novel Reference Genes in Acute Lymphoblastic Leukemia for Droplet Digital PCR
Genes doi: 10.3390/genes10050376
Authors:
Vanessa Villegas-Ruíz
Karina Olmos-Valdez
Kattia Alejandra Castro-López
Victoria Estefanía Saucedo-Tepanecatl
Josselen Carina Ramírez-Chiquito
Eleazar Israel Pérez-López
Isabel Medina-Vera
Sergio Juárez-Méndez
Droplet digital PCR is the most robust method for absolute nucleic acid quantification. However, RNA is a very versatile molecule and its abundance is tissue-dependent. RNA quantification is dependent on a reference control to estimate the abundance. Additionally, in cancer, many cellular processes are deregulated which consequently affects the gene expression profiles. In this work, we performed microarray data mining of different childhood cancers and healthy controls. We selected four genes that showed no gene expression variations (PSMB6, PGGT1B, UBQLN2 and UQCR2) and four classical reference genes (ACTB, GAPDH, RPL4 and RPS18). Gene expression was validated in 40 acute lymphoblastic leukemia samples by means of droplet digital PCR. We observed that PSMB6, PGGT1B, UBQLN2 and UQCR2 were expressed ~100 times less than ACTB, GAPDH, RPL4 and RPS18. However, we observed excellent correlations among the new reference genes (p < 0.0001). We propose that PSMB6, PGGT1B, UBQLN2 and UQCR2 are housekeeping genes with low expression in childhood cancer.
Source: Genes - Category: Genetics & Stem Cells Authors: Vanessa Villegas-Ru íz Karina Olmos-Valdez Kattia Alejandra Castro-L ópez Victoria Estefan ía Saucedo-Tepanecatl Josselen Carina Ram írez-Chiquito Eleazar Israel P érez-López Isabel Medina-Vera Sergio Ju árez-Méndez Tags: Article Source Type: research
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