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Use The Manual Assay As The Standard Reference And Trouble Shoot Each Step Of The Protocol Essay

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use the manual assay as the standard reference and trouble shoot each step of the protocol to improve the design of the automated assay. We will also test additional reagents such as protease K, hybridization mix, and wash buffers in order to improve the robustness of the protocol.

Aim-3: To validate the automated mRNA FISH assay of ARv7, ETS gene fusion and PTEN on DeNovo’s JETTA™ microfluidic chip, Jetta 400 sample preparation system and Vanguard image processing system. Demonstrate the entire system capable of automated single cell mRNA FISH on CTCs using blood samples from patients with metastatic prostate cancer.
This objective consists of two tasks as follows: Task 3.1: Patient blood sample collection and processing. We will collect blood samples from metastatic cancer patients before and after treatment. Dr. Clara Hwang, hematologist-oncologist and Dr. Mani Menon, Urologic surgeon, from the Henry Ford Health System will coordinate the efforts to select the patient population for the study. Letter of support from Drs. Menon and Hwang included. We propose to test 50 from African American patients and 50 samples from Caucasian during the study period. Samples prior to and after treatment will be collected for each patient and processed using the instrument pipeline set up for this study. We will correlate the marker status identified from CTC with the primary biopsy or prostatectomy samples using standard IHC, FISH and RNA ISH on FFPE tissues to validate our finding in

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