Screening for a set of recurrent mutations

Question 1

___ may be carried out with primers known as ___, which are specific at their 3’ end for either the mutant or the normal sequence.

Answer 1

allele-specific PCR; allele specific oligonucleotides

Question 2

The principle of this method is that PCR amplification takes place only when the 3 ′ nucleotide of the primer can correctly base - pair with the template DNA.

Answer 2

allele-specific PCR such as in amplification refractory mutation system or ARMS test

Question 2

Method for the detection of pre-defined mutations is the ____, in which the allele - specific primers for different mutations possess 5′ tails of different lengths.

Answer 2

oligonucleotide ligation assay (OLA)

Question 3

In ___ method, the 3 ′ end of a correctly base - paired oligonucleotide is able to be ligated to an adjacent common fluorescent primer and the ligated product can then be identified on a DNA sequencer.

Answer 3

oligonucleotide ligation assay (OLA)

Question 4

In Oligonucleotide Ligation Assay, several mutations can be tested simultaneously because the ____ permits the ligation products to be distinguished subsequently by their length.

Answer 4

differing 5’ tail lengths

Question 5

Methods based on ARMS and OLA have been used successfully in diagnostic laboratories for the detection of many common ___

Answer 5

Cystic fibrosis gene mutations

Question 5

It is possible to identify a mutation by the ____, using PCR to amplify the region followed by an appropriate restriction digest and subsequent gel electrophoresis to check the products.

Answer 5

loss or gain of a specific restriction site

Question 6

____ can, of course, be used to detect a predefined mutation although this may be less efficient when several samples are to be checked for the same set of specific base changes located in different exons.

Answer 6

DNA sequencing