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| AN IMPROVED TECHNIQUE FOR THE INSITU DETECTION OF DNA AFTER POLYMERASE CHAIN-REACTION AMPLIFICATION |
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| Author(s): NUOVO GJ, GALLERY F, MACCONNELL P, BECKER J, BLOCH W |
| Source: AMERICAN JOURNAL OF PATHOLOGY Volume: 139 Issue: 6 Pages: 1239-1244 Published: DEC 1991 |
| Times Cited: 168 References: 5 |
| Abstract: In situ detection of polymerase chain reaction (PCR)-amplified DNA in cell and tissue preparations previously required 5 to 7 primer pairs designed to generate a long (> 1,000 base pair) product. The authors describe a nonisotopic PCR in situ technique, employing a single primer pair and target sequences as short as 115 base pairs, that can detect one target molecule per cell. The essential procedural change is to withhold the DNA polymerase or primers until the reaction temperature approaches 80-degrees-C. The Hot-Start method greatly increased amplification specificity which, more than product size, appears to determine success of in situ PCR. The marked improvement in specificity may permit target detection by direct incorporation of labeled nucleotides. |
| Document Type: Note |
| Language: English |
| Reprint Address: NUOVO, GJ (reprint author), SUNY STONY BROOK, DEPT PATHOL, STONY BROOK, NY 11794 USA |
Addresses:
1. CETUS CORP, EMERYVILLE, CA 94608 USA
2. SUNY STONY BROOK, DEPT OBSTET & GYNECOL, STONY BROOK, NY 11794 USA |
| Publisher: AMER SOC INVESTIGATIVE PATHOLOGY, INC, 428 EAST PRESTON ST, BALTIMORE, MD 21202-3993 |
| Subject Category: Pathology |
| IDS Number: GV794 |
| ISSN: 0002-9440 |
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| | | | | | | | | Record from Web of Science® | | | | | | | | | |