Abstract
Mutations consisting of internal tandem duplications (ITDs) in exons 11 and 12 of the protooncogene c-kit are found in 30-50% of malignant canine mast cell tumors (MCTs). Traditionally, identification of such mutations in tumor specimens has been undertaken using standard polymerase chain reaction (PCR) and agarose gel electrophoresis. This procedure is limited to the detection of insertions and deletions larger than 9 base pairs in size. The purpose of this study was to compare the efficiency and accuracy of standard agarose gel electrophoresis with fluorescent polyacrylamide gel electrophoresis (PAGE) for the detection of ITDs in canine MCTs. The results of this study demonstrate that PAGE of labeled PCR products accurately predicts the size of the ITD in each tumor. In addition, other small insertions and deletions were not identified, suggesting that if they occur in canine MCTs, they do so infrequently. Because fluorescent and polyacrylamide formats are automated and have better resolution than agarose gels, fluorescent PAGE provides a more accurate, economical, and higher throughput method for the detection of c-kit mutations in canine MCTs.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 95-100 |
| Number of pages | 6 |
| Journal | Journal of Veterinary Diagnostic Investigation |
| Volume | 16 |
| Issue number | 2 |
| State | Published - Mar 2004 |
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ASJC Scopus subject areas
- Microbiology
- veterinary(all)
Cite this
Detection of c-kit mutations in canine mast cell tumors using fluorescent polyacrylamide gel electrophoresis. / Jones, Cameron L R; Grahn, Robert A; Chien, May B.; Lyons, Leslie A; London, Cheryl A.
In: Journal of Veterinary Diagnostic Investigation, Vol. 16, No. 2, 03.2004, p. 95-100.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of c-kit mutations in canine mast cell tumors using fluorescent polyacrylamide gel electrophoresis
AU - Jones, Cameron L R
AU - Grahn, Robert A
AU - Chien, May B.
AU - Lyons, Leslie A
AU - London, Cheryl A.
PY - 2004/3
Y1 - 2004/3
N2 - Mutations consisting of internal tandem duplications (ITDs) in exons 11 and 12 of the protooncogene c-kit are found in 30-50% of malignant canine mast cell tumors (MCTs). Traditionally, identification of such mutations in tumor specimens has been undertaken using standard polymerase chain reaction (PCR) and agarose gel electrophoresis. This procedure is limited to the detection of insertions and deletions larger than 9 base pairs in size. The purpose of this study was to compare the efficiency and accuracy of standard agarose gel electrophoresis with fluorescent polyacrylamide gel electrophoresis (PAGE) for the detection of ITDs in canine MCTs. The results of this study demonstrate that PAGE of labeled PCR products accurately predicts the size of the ITD in each tumor. In addition, other small insertions and deletions were not identified, suggesting that if they occur in canine MCTs, they do so infrequently. Because fluorescent and polyacrylamide formats are automated and have better resolution than agarose gels, fluorescent PAGE provides a more accurate, economical, and higher throughput method for the detection of c-kit mutations in canine MCTs.
AB - Mutations consisting of internal tandem duplications (ITDs) in exons 11 and 12 of the protooncogene c-kit are found in 30-50% of malignant canine mast cell tumors (MCTs). Traditionally, identification of such mutations in tumor specimens has been undertaken using standard polymerase chain reaction (PCR) and agarose gel electrophoresis. This procedure is limited to the detection of insertions and deletions larger than 9 base pairs in size. The purpose of this study was to compare the efficiency and accuracy of standard agarose gel electrophoresis with fluorescent polyacrylamide gel electrophoresis (PAGE) for the detection of ITDs in canine MCTs. The results of this study demonstrate that PAGE of labeled PCR products accurately predicts the size of the ITD in each tumor. In addition, other small insertions and deletions were not identified, suggesting that if they occur in canine MCTs, they do so infrequently. Because fluorescent and polyacrylamide formats are automated and have better resolution than agarose gels, fluorescent PAGE provides a more accurate, economical, and higher throughput method for the detection of c-kit mutations in canine MCTs.
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M3 - Article
VL - 16
SP - 95
EP - 100
JO - Journal of Veterinary Diagnostic Investigation
JF - Journal of Veterinary Diagnostic Investigation
SN - 1040-6387
IS - 2
ER -