Atlas of Hematology

Cloning of chimerical translocations as positive control for molecular genetic diagnosis of leukemia
The diagnosis of leukemia-specific mRNAs by polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR) require well-known positive standard controls. In general, the positive controls are obtained from cell lines and leukemia patients who have been diagnosed at the molecular level by RT-PCR. These are expensive and restricted sources for standard positive controls. Thus, there is a need for less expensive and reproducible standard positive controls in this area. We have cloned the t (9: 22) p190, t (9: 22) p210, t (4: 11), t (1: 19), t (15: 17), t (12; 21) breakpoint junctions of fusion genes into the plasmids. Cloned fusion genes are suitable for testing PCR experiments of the molecular genetic diagnosis of leukemia samples. We cloned and optimized fusion gene junctions as a standard positive control to check PCR efficiency and as a standard positive marker for diagnosis.

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Authors: Duran Ustek, Sema Sırma, Aris Cakiris, Fulya Coşan, Başar Oku, Uğur Özbek

Keywords: Füzyon genler, klonlama, ters transkriptaz PCR

Year: 2008
Number: 1
Referance No: 25349

Figure 1. Ethidium bromide stained agarose gel showing PCR from cloned fusion fragments. a) M-BCR-ABL, b)m- BCR-ABL, c) MLL-AF4, d)E2A-PB:1, e) PML-PARA variants, and f) t(12,21)