| First Author | Vinyals A | Year | 1998 |
| Journal | Clin Exp Metastasis | Volume | 16 |
| Issue | 7 | Pages | 603-11 |
| PubMed ID | 9932607 | Mgi Jnum | J:53667 |
| Mgi Id | MGI:1333267 | Doi | 10.1023/a:1006584910365 |
| Citation | Vinyals A, et al. (1998) Detection of differentially expressed gelatinase A in metastatic and non-metastatic subpopulations of tumor cells by target RNA arbitrarily primed polymerase chain reaction (TRAP-PCR). Clin Exp Metastasis 16(7):603-11 |
| abstractText | We have developed a novel procedure called Targeted RNA AP-PCR (TRAP- PCR) to quantitatively measure specific mRNA expression. The target mRNA is reverse transcribed using a specific primer and PCR is performed under low stringency conditions to generate a rich fingerprint-type band pattern. In this situation multiple sequences are coamplified with the targeted sequence. The amplification is carried out in a competitive fashion and is, in consequence, quantitative. We have applied this technique to determine Gelatinase A (Gel A) mRNA expression in the MXT mouse mammary carcinoma system. TRAP-PCR analysis using primers for Gel A produced a reproducible fingerprint including one major band whose identity was confirmed to be Gel A cDNA. Highly metastatic MXT subclones show an increased Gel A expression. Results were confirmed by Northern blot and protein activity (gelatin zymography). TRAP-PCR is a simple, sensitive and specific technique to comparatively quantify mRNA expression and requires less template than conventional methods. |
