In Situ
Hybridization Data Sheet
Cross-reference to notes or
photographs of slides or materials:
Master-mix:
Component |
Final
concentration/amount in hybridization mix |
Amount
for 1 slide |
Multiply
by number of slides +1 |
100%
Formamide |
50% |
20
µl |
|
20x
SSC |
2x |
4
µl |
|
50%
Dextran sulphate |
10% |
8
µl |
|
Salmon
sperm DNA 1µg/µl |
1µg |
1
µl |
|
10
mM EDTA |
0.125
mM |
0.5µl |
|
10%
SDS |
0.125% |
0.5µl |
|
Total |
|
34µl |
|
Component
|
Typical
value per slide |
Slide
codes and details
|
|
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
Master-mix
|
34
µl
|
|
|
|
|
|
|
|
|
|
|
Probe
1 |
25-200ng
|
|
|
|
|
|
|
|
|
|
|
Probe
2 |
25-200ng
|
|
|
|
|
|
|
|
|
|
|
Probe
3 |
25-200ng
|
|
|
|
|
|
|
|
|
|
|
Probe
4 |
25-200ng
|
|
|
|
|
|
|
|
|
|
|
Probe
5 |
25-200ng
|
|
|
|
|
|
|
|
|
|
|
Probe
6 |
25-200ng
|
|
|
|
|
|
|
|
|
|
|
Blocking
DNA |
10-100x
of probe
|
|
|
|
|
|
|
|
|
|
|
Water
|
(to
40µl)
|
|
|
|
|
|
|
|
|
|
|
Total
Volume
|
40
µl
|
|
|
|
|
|
|
|
|
|
|
Probe details
|
Probe
|
Code
|
Label
|
Labelling
method
|
Date
|
By
whom
|
1 |
|
|
|
|
|
|
2 |
|
|
|
|
|
|
3 |
|
|
|
|
|
|
4 |
|
|
|
|
|
|
5 |
|
|
|
|
|
|
6 |
|
|
|
|
|
|
Denaturation times
and temperatures for the hybridization mixture:
Denaturation times
and temperatures for the chromosome preparations:
Hybridization
start: |
finish:
|
time: |
Hybridization temperature:
Stringent washing
conditions:
SSC |
Formamide |
Temperature
°C |
Pat Heslop-Harrison
& Trude Schwarzacher Jan 2004
|