4 .5 kb long cDNA is used to identify a 250-kb BAC clone from a genomic library that fully contains CFTR gene. a. Question asked to describe the approaches that can be used to sequence each of these clones. b. Assume that there is a high possibility of diseasecausing mutations in this gene which are within exons or at intron–exon boundaries. If this assumption is correct, question asked to outline the method to identify the mutations in patients by using least amount of sequencing. Introduction: CFTR channels are present on epithelial cells of various organs (livers, lungs, pancreas, digestive track, reproductive track); it maintains water and salt balance over the surface of the epithelial cell. Due to mutation in CFTR, mucous accumulates and hardens the surface of the epithelial cell. Cystic fibrosis transmembrane conductance regulator(CFTR) is the transmembrane Cl - (Chlorine) channel. The gene of CFTR is present on chromosome 7 . Mutations that occur in the exon region of gene can be found by using different sequencing methods. Sequencing of cDNA is easier than sequencing of complete DNA, because the number of fragments is less.

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Chapter 16, Problem 16P

Summary Introduction

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4.5 kb long cDNA is used to identify a 250-kb BAC clone from a genomic library that fully contains CFTR gene.

a. Question asked to describe the approaches that can be used to sequence each of these clones.

b. Assume that there is a high possibility of diseasecausing mutations in this gene which are within exons or at intron–exon boundaries. If this assumption is correct, question asked to outline the method to identify the mutations in patients by using least amount of sequencing.

Introduction:

CFTR channels are present on epithelial cells of various organs (livers, lungs, pancreas, digestive track, reproductive track); it maintains water and salt balance over the surface of the epithelial cell. Due to mutation in CFTR, mucous accumulates and hardens the surface of the epithelial cell. Cystic fibrosis transmembrane conductance regulator(CFTR) is the transmembrane Cl^- (Chlorine) channel. The gene of CFTR is present on chromosome 7. Mutations that occur in the exon region of gene can be found by using different sequencing methods. Sequencing of cDNA is easier than sequencing of complete DNA, because the number of fragments is less.

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