MS-MLPA

Question 1

MS-MLPA

Answer 1

• semi-quantitative method to determine whether template DNA is methylated
• MS-MPLA probes are designed to hybridize to an area on the target DNA that includes a CpG methylation restriction enzyme site

Question 2

DMNT

Answer 2

• family of DNA methyltransferases
• catalyzes the transfer of a methyl group from S-adenyl methionine to the fifth carbon of cytosine residue = forms 5-methylcytosine

Question 3

Transcription is ultimately regulated by the interaction of multiple __ mechanisms that cooperate to __ or __ gene expression

Answer 3

Transcription is ultimately regulated by the interaction of multiple EPIGENETIC mechanisms that cooperate to ACTIVATE or SILENCE gene expression

Question 4

Methylation is regulated by proteins such as __ and__ that are involved in the active addition or chemical modification of DNA methylation

Answer 4

Methylation is regulated by proteins such as DMNT and Tet (purple) that are involved in the active addition or chemical modification of DNA methylation.

Question 5

How do DMNTs suppress gene expression ?

Answer 5

DMNT targets CpG sites and actively methylates DNA

Question 6

Hha1

Answer 6

• a restriction endocnuclease*
• recognizes GCGC sequence
• activity is blocked by methylation of cytosine of the first C

*NOTE: cuts at the middle of DNA segments, not at ends

Question 7

WIP Desaturation

Answer 7

98 degrees C for 5 min
GCGC Hha1 rec

Question 8

MS-MLPA Tube 2

Answer 8

• Ligation & Hha1 Digestions
• Unmethylated and methylated strands get ligated
• Unmethylated GCGC strand is cleaved
• Methylated GCGC are protected from cleavage

Question 9

WIP Multiplex Ligation-dependent Probe Amplification

Question 10

Products from Multiplex Ligation-dependent Probe Amplification are measured using __

Answer 10

Products from Multiplex Ligation-dependent Probe Amplification are measured using CAPILLARY ELECTROPHORESIS

Question 11

T or F: MS-MLPA can be used to identify Angelman and Prader-Willi Syndrome