Lecture 19 Part 1 - MS-MLPA

Question 1

What is MS-MLPA

Answer 1

a semi-quantitative method for determining the methylation status of template DNA

Question 2

How do the probes work in MS-MLPA

Answer 2

they are designed to hybridize to an area on the target DNA that includes a CpG methylation restriction enzyme site

Question 3

What is Hhal

Answer 3

a restriction endonuclease that recognizes GCGC sequences blocked by the methylation of cytosine of the first C

Question 4

What are the probes for MS-MLPA

Answer 4

same as MLPA with the additions of a Hhal recognition sequence that binds to the Hhal site

Question 5

What are the steps of MS-MLPA

Answer 5

denaturation and hybridization occur, then it is split into two tubes; one tube is digested, and the other is not. Then, both tubes are ligated and undergo PCR and capillary electrophoresis before being analyzed

Question 6

How does methylation effect digestion

Answer 6

if it is methylated it is not digested, if it is methylated it is digested and there is no PCR product