From the conception of the PRINS to
its coronation
Régen DROUIN, Geneticist
MD, PhD, FACMG, FCCMG
Department of Medical Genetics, CHUS
& Department de Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Cytogenetics:
- Chromosome Cytogenetics
- Interphase Cytogenetics
- Conventional Cytogenetics
- Molecular Cytogenetics
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Molecular Cytogenetic
Techniques available:
- FISH (Fluorescence In Situ Hybridization)
& variants: Q-FISH, express FISH, etc.
- PRINS (PRimed IN Situ labeling)
- M-FISH (Multicolor-FISH) or SKY
(spectral Karyotype)
- Band-FISH
- CGH (Comparative Genomic Hybridization)
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
PRINS
PRimed IN Situ
labeling
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
PRINS: Beginning
 PRINS technique was introduced in 1989 by Koch & al.Dept of Cytogenetics, Société Danoise du CancerAarhus, Danemark
 Ref. Koch J, Kolvraa S, Petersen K, Gregersen N, Bolund
L, Oligonucleotide-priming methods for the chromosomespecific labelling of alpha satellite DNA In
Situ.Chromosoma 1989;98:259-65
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
PRINS: HISTORY
 Method described 15 years ago, that was applied to
respond to questions regarding the structure of the minute
chromosomes in the primate -satellite DNA sequence.
 Development and Applications of PRINS (many variants
of the technique have been described)
 There are more and more targets investigated using
PRINS (many species: animals and plants)
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
 Multi-PRINS and Blocking step,
 Dual-color PRINS without blocking step
 New strategy for triple-color PRINS
 Nucleotids labeled with Bio-dUTP & Dig-dUTP
 Omission of blocking step
 Creating new color by mixing colors
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Extended section
of chromosome
Chromatin fiber of
packed nucleosomes‘String-of-beads’ DNA double helix
form of chromatin
Condensed section
of chromosome
10nm
700nm
30nm
300nm
1 400nm
2 nm
DNA
Condensation
and fiberFISH
A good PRINS method
should have:
- An extremely high specificity
(extremely low background)
- A good sensitivity (good
hybridization efficiency)
- Unambiguous recognition of the
hybridization signal
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
PRINS
Annealing
Elongation
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
MB
PRINS targets :
-
Metaphase Chromosomes
Interphase Nuclei
Fixed Tissues
Cells in culture
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
METHOD
Chromosome banding
• Slide preparation and the thermocycler
• Add Taq polymerase to the reaction solution
• Put the mix solution on the slide + cover slip
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
METHOD
Add Taq polymerase to the reaction solution
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
METHOD
Put the mix solution on the slide + cover slip
support thermocycler
METHOD
Detection & visualisation
• Washings post-PRINS
• Detection using a labeled antibody
• Washing of the antibody + counterstaining
• Observation under a fluorescence microscope
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Applications of the PRINS
technique:
- Identification of chromosomes
- Aneuploidy detection
- Analysis centromere DNA
- Identification of markers
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Proportions (%) of nuclei carrying
two signals of chromosome 8
Comparison of PRINS with FISH in the detection of
interphase nuclei carrying two signals of
chromosome 8 in 16 control cases
120
100
80
60
PRINS 8
FISH 8
40
20
0
1
2
3
4
5
6
7
8
9
Cases
10 11 12 13 14 15 16
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Proportions (%) of nuclei carrying
two signals of chromosome 7
Comparison of PRINS with FISH in the detection
of interphase nuclei carrying two signals of
chromosome 7 in 16 control cases
120
100
80
60
PRINS 7
FISH 7
40
20
0
1
2
3
4
5
6
7
8
9
Cases
10
11
12
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
13
14
15
16
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
A
B
PRINS 7
FISH 8
PRINS 8
D
FISH 7
C
E
Dual-PRINS 7 (red) and 8 (green)
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
F
 The development of molecular cytogenetics such as
FISH and PRINS are especially important for the
study of hematologic disorders. With these
approaches, not only dividing cells, but also nondividing cells can be studied for chromosome
identification.
 PRINS technique is a good alternative to FISH to
identify chromosomes both in metaphase and in
interphase nuclei. Our results showed no significant
difference between these two techniques regarding
detection sensitivity and specificity. PRINS is more
cost-effective, easier and faster than FISH.
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
• Samples preparation: usual techniques
• PRINS reaction and detection solutions for three
chromosome targets:
System
Labeling-chromosome
1
Bio- 7 Dig- 8 Bio- 18
2
Dig- 7 Bio- 8 Dig- 18
Detection mix
Avidin-fluorescein/
Anti-dig-rhodamine
Anti-dig-fluorescein/
Avidin-rhodamine
• Triple-PRINS programming:
1st cycle: 62.5C, 10 min with first label, wash in PBS 2 min
2nd cycle: 62.5C, 10 min with second label, wash in PBS 2 min
3rd cycle: 62.5C, 10 min with third label, wash in wash buffer 2 min
• Detection, Counterstain and image analysis
The results of triple-PRINS using two different labeling
orders combined with two different detection systems
Detection system 1a
Detection system 2b
Labeling order
1st signal 2nd signal 3rd signal
1. bio-dig-bio
2. dig-bio-dig
Yellow
Red
ambiguous
1st signal 2nd signal 3rd signal
Green
ambiguous
Yellow
Red
a: Mix of avidin-fluorescein/anti-dig-rhodamine;
b: Mix of anti-dig-fluorescein/avidin-rhodamine.
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Green
Primers
Name Chromosome
Sequence
Reference
-7
7
GCTTGAAATCTCCACCTGAAATGCCACAGC Koch et al. 1995
8c
18c
8
18
CTATCAATAGAAATGTTCAGCACAGTT
ATGTGTGTCCTCAACTAAAG
Pellestor et al. 1996
Pellestor et al. 1995
Xc
X
GTTCAGCTCTGTGAGTGAAA
Pellestor et al. 1995
D599
D600
Y
Y
TGGGCTGGAATGGAAAGGAATCGAAAC
TCCATTCGATTCCATTTTTTTCGAGAA
Speel et al. 1995
Speel et al. 1995
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
A: bio-dig-bio labeling order with detection system 1
The first PRINS reaction
3’
5’
The second PRINS reaction
3’
5’
3’
5’
The third PRINS reaction
3’
5’
3’
3’
3’
5’
: Chromosome DNA;
: The first primer;
: dNTP;
The second PRINS site
on chromosome 8
The first PRINS site on
chromosome 7
The third PRINS site on
chromosome 18
5’
The second PRINS site
on chromosome 8
5’
The first PRINS site on
chromosome 7
: Biotin-dUTP;
: The second primer;
The first PRINS site on
chromosome 7
: Digoxigenin-dUTP;
: The third primer;
B: dig-bio-dig labeling order with detection system 2
The first PRINS reaction
3’
5’
The first PRINS site on
chromosome 7
5’
The second PRINS site
on chromosome 8
5’
The first PRINS site on
chromosome 7
The second PRINS reaction
3’
3’
The third PRINS reaction
3’
5’
3’
3’
3’
5’
: Chromosome DNA;
: The first primer;
: dNTP;
5’
The second PRINS site
on chromosome 8
5’
The first PRINS site on
chromosome 7
: Biotin-dUTP;
: The second primer;
The third PRINS site on
chromosome 18
: Digoxigenin-dUTP;
: The third primer.
bio-dig-bio labeling,
avidin-fluorescein/ Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
anti-dig-rhodamin detection
bio-dig-bio labeling,
avidin-fluorescein/
anti-dig-rhodamin detection
bio-dig-bio labeling,
avidin-fluorescein/ Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
anti-dig-rhodamin detection
dig-bio-dig labeling,
anti-dig-fluorescein/Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
avidin-rhodamin detection
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
XY normal
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<==Multi-PRINS
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FISH==>
47,XX,+18
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<==Multi-PRINS
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FISH==>
Main advantages of the PRINS technique:






Semi-automated Protocole because of the use of thermocycler
Simple, reproducible and reliable method
Very good ratio cost benefit
Short duration of the reaction (on the average)
Availability of any primers (automatic synthesizer)
The specificity and small size of the primers (18 à 35 mer) does not
generate cross reaction
 Specificity of centromeric sequences of chromosomes 13 and 21
 More efficient in some cells with more condensed nuclei: the size of
primer is much smaller than the size of the probe thus making it
easier to pass the nuclear membrane and hybridize to the target DNA
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Main advantages of the PRINS technique:
 Detection of unique intragenic sequences
 Very low background because of the absence of labeling directly on
the primer and the rapidity of the reaction
 Good preservation of the integrity of the chromosome structure due to
the short incubation time
 Multi-target detection: it is very easy to do multi-color PRINS or
combine with FISH to simultaneously detect different chromosomes
in the same cells.
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
CONCLUSIONS
Technique PRINS
Powerful technique, simple and universal
Represents a very good alternative to FISH
Future Perspectives
Detection of fusion genes
Detection of female foetal cells using the
technique of PRINS-RNA
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Applications of the FISH
and PRINS techniques:
- Identification of chromosomes
- Détection of aneuploïdies
- Analysis of centromeric DNA
- Identification of marker chromosomes
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
A good PRINS method
should have:
- An extremely high specificity
(extremely low background)
- A good sensitivity (good
hybridization efficiency)
- Unambiguous recognition of the
hybridization signal
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Same nucleus in both panels is shown with the opposite colours
Simultaneous FISH analysis using sex chromosome Simultaneous reverse FISH analysis using
specific probes: the red signal correspond to the
sex chromosome specific probes: the red
X chromosome and the blue signal to the Y
signal correspond to the Y chromosome and
chromosome
the blue signal to the X chromosome
FISH technique (LSI-21 probe) combined
to PRINS reaction (Y primer sequence)
Male fetal cell with
3 copies of Chromosome 21 (red signals) and
1 copy of the Y chromosome (green signal)
PRINS technique
Qu ickTi me ™ a nd a TIFF (Un co mpre ssed ) d eco mp re ssor are n ee de d to se e thi s pi cture .
Male fetal cell with trisomy 18
Fetal cell with trisomy 18
detected by PRINS technique
Fetal triploid cell detected by PRINS technique
using centromeric
PRINS (primer
oligonucleotide
centromeric oligonucleotide
primer specific
8 to the
chromosome 7Fetal
(the cell
nucleus
with with
triploidy
three red signals)
PRINS technique
Triploidic fetal cell with 3 copies of chromosome 7
FETAL CELLS PROJECT: QUANTIFICATION OF ALL F ETAL
NUCLEATED CELLS FROM PERIPHERAL BLOOD OF PREGNANT
WOMEN BEARING ANEUPLOID CONCEPTUS USING MOLECULAR
CYTOGENETIC TECHNIQUES (FISH AND PRINS TECHNIQUES)
Table II: Clinical data of the patients and the number of fetal cells
per mL of maternal blood in 12 cases of male trisomy 21
Patient
No.
Number Number
Pare ntal Grav idity Indi cation for
Week of
Parity
Family history
of
of fetal
age
amniocentesis*
gestation
slides/ml cells/ml
Abortion
Mat. Pat.
1
42 41
G4P1A2
AMA
Hypo thyroidy
18 3/7
42
32
2
32 35
G3P1A1
Antecedents
T21 / S B
18
42
24
3
34 32
G2P0A1
Antecedents
Cardiac defects
19
45
11
4
26 27
G2P1A0
NT
Pyloric steno sis
17 3/7
31
8
5
36 39
G5P3A1
AMA
47,XXY
19 4/7
27
10
6
37 41 G3P2A0
AMA / NT
C. V. Pb
21
31
14
7
35 32 G5 P4A0
AMA
18 2/7
24
10
8
42 43 G5P2A2
AMA
Nil
15 4/7
25
11
9
39 34 G1P0A0
AMA
SB / Others
20
20
8
10
40 37 G4P1A2
AMA
Diabetes /
Cance r
17 1/2
27
6
11
38 36 G2P1A0
AMA
Thyroiditis / Cri
du chat
13
19
9
AVC /
Hypercholeste
rol.
Paternal
20
7
18
Ysat
Legend: *AMA: Advanced maternal age; AVC: Atrio Ventricular Canal;
CV pb: Cardio Vascular
Problem ; CPC: Choroid Plexus Cyst; NT: Nuchal
translus cency; SB: Spina Bifida.
12
26 33 G1P0A0
NT
Table III: Clinical data of the patients and the number of
fetal cells per mL of maternal blood in 7 cases of
male trisomy 18
Parental
age
Gravidity
Indication for
Parity
amniocentesis*
Abortion
Family
history
Mat.
Pat.
41
?
G3P0A2
AMA / IUGR +
Multi ple Defects
Nil
38
33
G2P1A0
AMA / IUGR
Cardiac Defects
26
24
G1P0A0
Nil
28
29
G2P0A1
30
31
40
38
Week of
gestation
Number Number
of
of fetal
slides/ml cells/ml
3/7
26
11
17
23
20
Nil
21 3/7
26
7
Cystic Hyg roma
Crohn disease/
Diabetes
18
25
4
G2 P1A0
Omphalocele/
strawberry-like head
Nil
17
2/7
18
12
?
G3 P1A1
AMA
Nil
17
6/7
20
4
?
G1P0A0
AMA / TT + / Cardiac
malformations
Twin pregn +
Multiple Sclerosis
22
5/7
20
4
18
Legend: *AMA: Advanced Maternal Age; IUGR: Intra-Uterin e Growth
Retardation;
TT+: Triple test +
Table IV: Clinical data of the patients and the number of fetal cells
per mL of maternal blood in 7 different other cases of
aneuploidies
Different
aneuploidies
Parental
age
Gravidity
Indication for
Parity
amniocentesis*
Abortion
Family
history
Weeks of
gestation
Number Number
of
of fetal
slides/ml cells/ml
Mat.
Pat.
69,XXX
27
29
G2P1A0
IUGR
C V pb
19
21
25
69,XXX
27
29
G1P0A0
2 ombili cal
vessels
Cance r + CV pb
21 5/7
20
10
47,XXX
38
35
G3P2A0
AMA
Cancer
3/7
18
16
47,XXY
32
31
G3P1A1
TT= 1/146
Diabetes
20
22
19
47,XYY
38
37
G7P2A4
AMA
HTA
17
26
26
46,XY,r(22)
39
38
G4 P2A1
Congenital defects
mat 16qh+
pat 9 peri inv
27
39
10
47,XY,+13
161/2
?
G1P0A0
CL + CD
Nil
24
20
9
Legend: *AMA: Advanced Maternal Age; CD: Cardiac Defect; CL: Cleft lip;
IUGR: Intra Uterin e
Growth Retardation; TT: Triple test; OPD: Oculo-Ph aryngeal
Dystrophy
21
CONCLUSIONS
 It is possible to detect fetal cells in every pregnant woman during
the pregnancy. Using molecular cytogenetic approaches, (FISH and
PRINS), we showed that there were between 2 and 6 fetal cells per
mL of maternal blood
 The low fluctuation between the 12 pregnant women we studied
is likely due to personal characteristics specific to each of the
women.
 The number of fetal cells in the maternal blood does not appear
to be influenced by previous pregnancies.
CONCLUSION
 It is possible to detect fetal cells between millions of maternal blood cells (our
previous results : 2 to 6 fetal cells per mL of maternal blood, see ref.3) using
molecular cytogenetic techniques (FISH et PRINS)
 In agreement with several reports, our preliminary results obtained with 14
pregnant women show a 5-fold higher number of fetal cells in pregnancies associated
with aneuploid conceptuses (see Tables 1, 2 & 3)
 This finding provides an interesting perspective for the development of a safe and
convenient non-invasive molecular cytogenetic prenatal diagnosis for the most
common fetal chromosomal aneuploidies (13, 18, 21, X and Y).
 However, this procedure is time consuming and labour intensive. More studies are
needed to confirm the robustness of this methodology and automation will be
required before widespread application.
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Telomere
Simple DNA sequence (T2AG3) tandemly repeated, of variable
length, located at the extremities of the chromosomes.
Telomeres are essential elements that protect the extremities of
the chromosomes from degradation and ligation.
Shortening
Incomplete Replication
Nuclease Activity
Equilibrium
Elongation
Addition of repetitions
T2AG3 by the telomerase
Senescence
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Telomeres
• Specialized structures made of DNA and PROTEINS
• Repeated DNA sequence: 2 à 15 kb
• Maintain the chromosome stability
TTAGGG TTAGGG TTAGGG
AATCCC AATCCC AATCCC
• Around 30 to 120 bp are lost per somatic cell division
• Too short : cellular senescence and genetic instability
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Measurement of telomeres
Average length of telomeres :
• Measurement of terminal restriction fragments.
– Digestion using restriction enzymes of purified DNA
– Visualization and measurements of telomeric fragments by
Southern blot
• Cleavage of telomeres at variable distance
• No information individual telomeres
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Measurement of telomeres
Length of individual telomeres :
Quantitative FISH (Q-FISH)
Hybridization telomeric PNA probes
Measurements of the signal intensity
Length Profil of individual telomeres
Variation of hybridization efficiency ???
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
The PRINS reaction
PRINS
Marquage par synthèse in situ amorcée
Appariement
The primer sequence is
complementary to the
telomeric sequence:
(CCCTAA)7
Élongation
Tissus
Koch et al., 1995
Terkelsen et al., 1995
Cellules
Chromosomes
Fibres
d’ADN
Figure 3:
Wrn ∆hel/ ∆hel
11-dUTP-digoxigénine
Department of Medical Genetics, CHUS & Department of Pediatrics,
62.5 °C, 10 minutes
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Le double-PRINS
• To increase the efficiency of the telomere labeling using
PRINS, Dr Ju Yan developed an innovative double-labeling
technique : two complementary primers, (CCCTAA)7 et
(TTAGGG)7, are used to label both DNA strands of the
telomere sequences.
: (TTAGGG)7
A
A
: (CCCTAA)7
C
B
B
C
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Materials & méthods
PRINS Reaction
1. The PRINS mixture is put on the slide :
dNTP, labeled dUTP (biotine or digoxigenine), primers, Taq buffer &
Taq polymerase
2. Hybridization and elongation :
20 minutes at 63°C,
on a thermocycler.
Materials & methods
Double-PRINS Procedure
1. 1st PRINS reaction : primer (CCCTAA)7.
2. Brief washings (washing buffer then PBS 1x ).
3. 2nd PRINS reaction : primers (TTAGGG)7.
4. Washings (washing buffer : 5 min. at 45oC and 2 x 5 min.
at room To).
5. Revelation of the signals : fluorescent antibodies.
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Materials & methods
Image Analysis
40 metaphases per case, 10 for each type of signals :
- green signals only
- red signals only
- double color signals
- double green signals
Analyzed using the ISIS 2 software of Metasystems
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Results
1. Comparison between simple PRINS and PNA-FISH
One PRINS cycle
PNA-FISH
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Results
2. Double-PRINS double color
Results
3. Comparison between simple PRINS and double-PRINS
Red only
Green only
Double color
Results
4. Comparison between double-green double-PRINS & PNA-FISH
Double-vert
PNA-FISH
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Results
Efficacité moyenne de marquage (%)
100
90
80
70
60
50
Vert
Rouge Double Double PNA
couleur vert
Vert
Rouge Double Double PNA
couleur vert
Marquage
des chromatides
bras des chromosomes
Department
of Medical Genetics, CHUSMarquage
& Departmentdes
of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Telomere signals on chromatin fibers (arrows) detected
by double-green labelling using PRINS.
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada
Fig. 4:
Single-copy sequence
detected by multi-primer
PRINS technique.
Locus:
AL672294, 37kb
on chromosome 1q
subtelomere, 28 primers.
Labeling: Biotin-dUTP
Detection: Avidin-FITC
Team of Dr Régen DROUIN
Cytogenetics
Molecular Genetics
• Walid DRIDI
Macoura GADJI
Kada KRABCHI
Josée LAVOIE
• Sandrine LACOSTE
Stéphane OUELLET
Patrick ROCHETTE
François VIGNEAULT
• Éric BOUCHARD
Marc BRONSARD
• Ju YAN
• Nathalie BASTIEN
Mélissa FERLAND
Isabelle PARADIS
Department of Medical Genetics, CHUS & Department of Pediatrics,
Université de Sherbrooke, Sherbrooke, Quebec, Canada