Balanced Translocation detected by FISH

1
Balanced Translocation detected by FISH
2
Red- Chrom. 5 probe
Green- Chrom. 8 probe
3
(No Transcript)
4
2D Protein Gels
5
(No Transcript)
6
MS-peptide size signature match to all predicted
proteins
7
X
2
3
Parent
X
Gamete
X
Child
8
A1
B1
C1
D1
X
2
3
E2
Parent
B2
C2
A2
A2
B1
C1
D2
X
Gamete
B1
C1
A2
X
Child
9
A1
B1
C1
D1
X
2
3
E2
Parent
B2
C2
A2
A2
B1
C1
D2
X
NR
Gamete
R
B1
C1
A2
X
Child
10
Positional Cloning by Recombination Mapping

• Follow the mutation
• 2. Follow which DNAs are
• co-inherited (linked)

11
Positional Cloning by Recombination Mapping

• Follow the mutation
• To determine disease gene
• presence or absence (genotype)
• from phenotype you must
• first establish
• Dominant / recessive
• Aurosomal / sex-linked

12
SINGLE GENE DEFECTS
Modes of Inheritance
To deduce who (likely) has one or two copies of
mutant gene
Affected Female
Unaffected Male
13
/
D/
D/
/
AUTOSOMAL DOMINANT
14
a/
a/
x/
/Y
x/
/Y
a/a
x/Y
RECESSIVE
RECESSIVE
X-LINKED
AUTOSOMAL
15
(No Transcript)
16
Positional Cloning by Recombination Mapping
2. Follow which DNAs are co-inherited
(linked) Use DNA sequences that differ among
individuals within a family- Polymorphisms.
17
VNTR / STRP DETECTION
18
(No Transcript)
19
A1
B1
C1
X
2
3
Parent
B2
C2
A2
A2
B1
C1
X
Gamete
B1
C1
A2
X
Child
20
Meiosis
Meiosis
Meiosis
A2 B2
A1 B1
21
Recombination Mapping
Measures distance between 2 sites on a chromosome
according to frequency of recombination
Distance between 2 DNA markers or Distance
between a disease gene and a DNA marker
22
No fixed proportional
Conversion between
Genetic distance (cM)
and
Physical distance (kb, Mb)
23
FAMILY A
24
FAMILY B
A1

A2
D
NR
NR
NR
NR
NR
R
R
R
R
R
R
NR
25
INFORMATIVE MEIOSIS
Ideally- unambiguous inheritance of mutation
and markers (requires heterozygosity for each
in parent) knowledge of which alleles linked in
parent (phase)
26
Assign numbers to results of linkage
analysis to deal with non-ideal meioses to
sum data from many meioses in a family to sum
data from several families
27
If unlinked-
?
If linked and RF
1/2
?
Likelihood of R
1 -
?
Likelihood of NR
1/2
Family A has 1 recombinant and 5 Non-Recombinants
?
Likelihood, given linkage of
Or given unlinked-
5
?
. (1- )
?
?
L ( )
6
L (1/2) (1/2)
28
Z 3
Lod
q
29
FAMILY B
A1

A2
D
NR
NR
NR
NR
NR
R
R
R
R
R
R
NR
30
Family B- Disease gene may be linked to A1 or A2
Consider equally likely
50 chance Family B has 1 R and 5 NR
50 chance Family B has 5 R and 1 NR
31
Phase known
Phase unknown
?
0.1 0.2 0.3 0.4 0.5
Z 0.28 0.32 0.22 0..08 0
32
For family A with meioses 1, 2, 3, 4 ..
Z Z1 Z2 Z3 Z4 ..
For multiple families, A, B, C, D..
Z Z(A) Z(B) Z(C) Z(D) .
Assumption same gene responsible for disease
in all families Problem locus heterogeneity
33
Z 3
Lod
q
34
(No Transcript)
35
LINKAGE DISEQUILIBRIUM
Many generations
36
PCR test DNA segments
37
SSCP
38
Normal
D
F506
39
(No Transcript)
40
Testing for specific mutations
41
ARMS 3 mis-match of primer
42
(No Transcript)
43
TaqMan
44
OLA
45
(No Transcript)
46
(No Transcript)
47
(No Transcript)
48
(No Transcript)