In Situ Hybridization Data Sheet

Date: ____ 20

 

RUN CODE: Name:

 


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
                   

 

Run: Date:

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