| Autor: |
Hakkı Ogun Sercan, Ilgın Öztürk, Ceyda Çalışkan, Melek Pehlivan, Zeynep Sercan |
| Jazyk: |
angličtina |
| Rok vydání: |
2012 |
| Předmět: |
|
| Zdroj: |
Turkish Journal of Hematology, Vol 29, Iss 4, Pp 348-353 (2012) |
| Druh dokumentu: |
article |
| ISSN: |
1308-5263 |
| DOI: |
10.5152/tjh.2011.76 |
| Popis: |
OBJECTIVE: At present in Turkey, centers that are able to give reliable RQ-PCR BCR-ABL results are limited in number. We aimed to describe a cost-effective, in-house method for BCR-ABL quantification and to illustrate an example for RQ-PCR validation tests. METHODS: BCR-ABL and ABL target sequences were cloned into pJET1.2 vectors; from which calibrators were prepared and used as templates in RQ- PCR reactions to generate standard curves. Dilutions of K562 cells (representing an in vitro simulation of BCR-ABL transcript reduction) were analyzed. RESULTS: Previously determined standard curves were used to calculate the BCR-ABL and ABL copy numbers. Linear BCR-ABL results were obtained. Repetitive experiments showed that our methodology is able to detect one BCR-ABL positive cell in a total of 1x105 cells. CONCLUSION: The approach described in this manuscript is suitable for implementation into any RQ-PCR instrument and/or kit aiming to quantify BCR-ABL transcripts. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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