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Table 5 (FISH guide)
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Table 5. Relative amounts of DNA probes and their corresponding amount of competitor DNA in FISH.
|
Probe type |
DNA probe amount |
Competitor DNA amount |
|
plasmid |
1-5 ng |
10-20 µg |
|
cosmid |
5-20 ng |
20-30 µg |
|
P1, BAC |
20-60 ng |
20-30 µg |
|
YAC (total yeast) |
50-100 ng |
20-40 µg |
|
YAC (Alu PCR) |
20-50 ng |
20-40 µg |
|
band/arm paint |
100-200 ng |
30-40µg |
|
chromosome paint |
100-500 ng |
30-40 µg |
|
M-FISH |
Several micrograms |
40-50 µg |
|
CGH (nick) |
500-2000 ng |
50-150 µg |
|
CGH (PCR) |
1000-2000 ng |
30-40 µg |
|
Repeat probe* |
1-5 ng |
none |
*A repeat probe (alpha, beta satellite or similar) can be used in the presence of any amount of competitor DNA but the amount of labeled probe taken has to be accordingly increased, otherwise the probe can be blocked out.
The amount of competitor DNA used is higher in simultaneous denaturing and lower in separate denaturing.