Methods for RNA extraction, cDNA preparation and analysis of CFTR transcripts.
The scope of this article is to outline some of the basic methods for good quality RNA preparation from mammalian tissues and cells (including epithelial cells). Additionally, we give an outline of common techniques of measuring CFTR gene expression such as quantitative and semi-quantitative reverse transcription (RT) PCR and ribonuclease protection assay (RPA). These methods are designed to detect low abundance transcripts, which apply to CFTR mRNA in most cell types and tissues.
Centre of Human Genetics, National Institute of Health, Lisboa, Portugal.
Anabela S
AS
Ramalho
Sebastian
S
Beck
Carlos M
CM
Farinha
Luka A
LA
Clarke
Ghanshyam D
GD
Heda
Bernhard
B
Steiner
Javier
J
Sanz
Sabina
S
Gallati
Margarida D
MD
Amaral
Ann
A
Harris
Maria
M
Tzetis
D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D005821:Genetic Techniques; D006801:Humans; D012313:RNA / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
C505032:CFTR protein, human; D018076:DNA, Complementary; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D012313:RNA