A Pipeline with Multiplex Reverse Transcription Polymerase Chain Reaction and Microarray for Screening of Chromosomal Translocations in Leukemia
The procedure of multiplex RT-PCR-microarray assay. (a) Steps of multiplex RT-PCR. Reverse transcription was performed on total RNA with multiple chimeric reverse primers. The resulting cDNA was amplified with two-round PCR. First round was amplified with multiple chimeric forward primers on the 5′ region (upstream of known translocation points) and universal primer SP6 on the 3′ region. Second round was with universal primer T7 and biotin labeled SP6, resulting in biotinylated complex. (b) Hybridization with microarray. Take the sample with AML1-ETO translocation as example.
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