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Table 1 (FISH guide)
TOPICS: |1| 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | 22 | 23
FIGURES: | 1 | 2 | 3 | 4 | 5 | 6 |7 | 8 | 9 | 10 | 11 |TABLES: | 1 | 2 | 3 | 4 | 5 | 6 | 7
Table 1.DNA probe labeling procedures.
NICK TRANSLATION | PCR LABELING |
1 ug DNA (20 ng/?l final conc.) 5 ?l (10x) nick translation buffer * 5 ?l (100mM=10x) ?-mercaptoethanol 2.5?l (1mM=20x) each dACG 5?l (0.325/0.175 mM=10x) dTTP/dUTP 5?l (1?g/?l=10x) DNase (Sigma), 1?l (10U/?l) E. Coli polymerase ultrapure sterilized water to 50 ?l. | Template DNA (various amounts) 2.5-3.5?l (10x) PCR buffer 0.15?l (33.3mM) each dACG 0.7?l (5mM) dTTP 1.6?l (1mM) dUTP 0.1-1?l (50?M) primer(s) 0.2-0.4?l (5U/?l) Taq polymerase ultrapure sterilized water to 25 ?l |
* = any E.coli polymerase buffer
dUTP = labeled dUTP deoxynucleotide.