b'Gel & Capillary | RUO AssaysGel and CapillaryPML/RAR t(9;22) Translocation Assays Research Use Only (RUO) AssayPML/RAR t(15;17) Translocation AssaysAssay Use Specimen RequirementsThis Research Use Only assay identifies PML/RAR t(15;17)This assay tests complementary DNA (cDNA) template.translocations.Summary and Explanation of the TestReferences1.H De Th et al., Nature 347: 558-561 (1990).Four master mixes are included in these assay kits. Master mixes2. H De Th et al., Cell 66: 675-684 (1991).are used to amplify complementary DNA (cDNA) produced from3. A Kakizuka et al., Cell 66: 663-674 (1991).specimen(s), as well as positive and negative RNA controls (included).4. WH Miller et al., Proc. Natl. Acad. Sci. 89: 2694-2698 (1992).Primers target an internal control transcript and the variety of Bcr1, Bcr2, and Bcr3 type transcripts expressed from PML-RAR translocations. Amplicon products can be analyzed by differential fluorescence detection using capillary electrophoresis or standard gel electrophoresis. A PML-RAR translocation is indicated if just one of the 2nd round master mixes (Mix 2b or Mix 2c) generates product(s) of the valid size. Reagents for RNA extraction and reverse transcription are not included. This assay is compatible with all standard RNA extraction and cDNA synthesis methods. This is a qualitative assay and has not been validated for quantitative use.Background Three PML/RAR translocation patterns have been identifiedin samples with acute myelogenous leukemia (AML): type A is theshort (S-form); the breakpoint occurs within breakpoint cluster region 3 (Bcr-3).Type B is the long (L-form); the breakpoint occurs within Bcr-1.There is a third type B variant or variable (V-form); the breakpoint is within Bcr-2.Identification of the PML/RAR t(15;17) rearrangements are commonly used in the study of APL becauseit is correlated with responsiveness to treatment.This RT-PCR method directly identifies the chimeric PML/RAR transcripts expressed from all three forms of PML/RAR translocations.Figure Legend: This figure shows the genomic organization of the PML andPML GeneRAR genes on chromosomes 15 and 17, respectively. Boxes represent exonChromosome 15regions of the PML (red boxes) and RAR (orange) encoding exons. The solid black line represents intron regions, which were left incompletely spliced toRARA Geneassist in demarcation of the exon segments. Primers are indicated by arrows,Chromosome 17and the size of several of the products are indicated below the translocated gene segments. S-form (Bcr3) and L-form (Bcr1) PML-RAR translocations are depicted in the lower portion of the figure. 124'