| Autor: |
Michelle A, Anderson, Dean E, Brenner, James M, Scheiman, Diane M, Simeone, Nalina, Singh, Matthew J, Sikora, Lili, Zhao, Amy N, Mertens, James M, Rae |
| Rok vydání: |
2010 |
| Předmět: |
|
| Zdroj: |
The Journal of molecular diagnostics : JMD. 12(5) |
| ISSN: |
1943-7811 |
| Popis: |
Background and aims: Biomarker use for pancreatic cancer diagnosis has been impaired by a lack of samples suitable for reliable quantitative RT-PCR (qRT-PCR). Fine needle aspirates (FNAs) from pancreatic masses were studied to define potential causes of RNA degradation and develop methods for accurately measuring gene expression. Methods: Samples from 32 patients were studied. RNA degradation was assessed by using a multiplex PCR assay for varying lengths of glyceraldehyde-3-phosphate dehydrogenase, and effects on qRT-PCR were determined by using a 150-bp and a 80-bp amplicon for RPS6. Potential causes of and methods to circumvent RNA degradation were studied by using FNAs from a pancreatic cancer xenograft. Results: RNA extracted from pancreatic mass FNAs was extensively degraded. Fragmentation was related to needle bore diameter and could not be overcome by alterations in aspiration technique. Multiplex PCR for glyceraldehyde-3-phosphate dehydrogenase could distinguish samples that were suitable for qRT-PCR. The use of short PCR amplicons ( |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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