Quantifying CFTR mRNA in Human and Mouse Islets

Total RNA from human and mouse islets were prepared as described [20 (link)]. CFTR mRNA expression was measured by RT-qPCR using primers and probes from Taqman mRNA assays (Life Technologies, California, USA). Human CFTR: Hs00357011_m1 expression was normalized against human HPRT1: 4333768 F, while mouse CFTR: Mm00445197_m1 expression was normalized against mouse HPRT1: Mm00446968. RT-qPCR runs were performed for individual batches of human (N = 5) and mouse (N = 5) islets in triplicate wells of 384-well plate on a 7900 HT RT-PCR system (Applied Biosystems, California, USA).

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Edlund A., Esguerra J.L., Wendt A., Flodström-Tullberg M, & Eliasson L. (2014). CFTR and Anoctamin 1 (ANO1) contribute to cAMP amplified exocytosis and insulin secretion in human and murine pancreatic beta-cells. BMC Medicine, 12, 87.

Publication 2014

Taqman mRNA assays 7900 HT RT-PCR system

Assays Cftr Human Mouse Mrna Primers Rt pcr

Corresponding Organization :

Other organizations : Lund University, Karolinska Institutet

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

CFTR mRNA expression in human islets
CFTR mRNA expression in mouse islets

control variables

Total RNA from human and mouse islets were prepared as described [20]
RT-qPCR runs were performed for individual batches of human (N = 5) and mouse (N = 5) islets in triplicate wells of 384-well plate on a 7900 HT RT-PCR system (Applied Biosystems, California, USA)
CFTR mRNA expression was normalized against human HPRT1: 4333768 F for human islets
CFTR mRNA expression was normalized against mouse HPRT1: Mm00446968 for mouse islets

controls

No positive or negative controls were explicitly mentioned in the protocol

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