Biomedical Engineering Reference
In-Depth Information
Female
DNA
(a)
Female
Tri 21
(b)
Figure 1.4
Detection of Down syndrome using the MLPA trisomy kit, including eight probes for
sequences on each of chromosomes 13, 18, 21 and X, respectively, in addition to three probes
for a chromosome Y target sequence. The arrows indicate the chromosome 21 probes. (a) The
electropherogram of an MLPA performed on normal female DNA. (b) MLPA performed on DNA
from a Down syndrome patient, clearly showing an increase of all chromosome 21 probes.
such as: Down's syndrome (see Figure 1.4), breast (
BRCA1
) [37] and colon cancer [32]
(
MSH2
and
MLH1
), Duchenne [36], cystic fibrosis, centrifugal muscular atrophy, sickle cell
anemia and Marfan syndrome, to name but a few. However, future MLPA kits will focus
more on the detection of aberrations of genes playing roles in tumorigenesis, apoptosis,
angiogenesis and pharmacogenetics.
PROTOCOL 1.9 Multiplex Ligation-Dependent Probe
Amplification on DNA
Equipment and reagents
Complete MLPA kit (MRC-Holland) containing:
—MLPA probe mix
—MLPA buffer
—Ligase-65 buffer A
—Ligase-65 buffer B
—Ligase-65
—ultra pure water
—10
×
SALSA PCR buffer
—10
×
SALSA PCR buffer
—SALSA PCR forward primer (FAM or D4-labeled): 5'-GGGTTCCCTAAGGGTTGGA-3'
—SALSA PCR reverse primer (unlabeled): 5'-GTGCCAGCAAGATCCAATCTAGA-3'
•
