1. Leukemia Cell Study Strode Note: Meaningless title.

2. Background Acute Lymphoblastic Leukemia (ALL): cancer of lymphocytes (white blood cells) Acute Lymphoblastic Leukemia (ALL): cancer of lymphocytes (white blood cells) –Accounts for 25% of childhood cancers, making it the most common malignancy in children less than 15 years of age. –Occurs most frequently at 2-5 years of age, and is more common in males and Caucasians. –Cure rates of ALL is approaching 80% percent. –Two different types of ALL are B-cell ALL and T-Cell ALL.  REH: type B-cell ALL cell line (immortalized cell)  Jurkat: type of T-cell ALL cell line. Strode Note: Too many words and no image.

3. Background Understanding how cancer occurs may lead to the development of better chemotherapy drugs, and perhaps even a biologically targeted therapy. Understanding how cancer occurs may lead to the development of better chemotherapy drugs, and perhaps even a biologically targeted therapy. Research is being done on the expression of Mer in Leukemia. Research is being done on the expression of Mer in Leukemia. Strode Note: Better but still no image.

4. Background Mer tyrosine kinase: a protein that has anti-apoptotic signaling (anti-death) providing a survival advantage to the cells and transforming capabilities allowing normal cells to turn into cancer cells. Mer is not normally expressed in lymphocytes. Mer tyrosine kinase: a protein that has anti-apoptotic signaling (anti-death) providing a survival advantage to the cells and transforming capabilities allowing normal cells to turn into cancer cells. Mer is not normally expressed in lymphocytes. –Mer is found in several cancers such as:  Gastric cancer  Pituitary adenoma  Mantle cell lymphoma  Alveolar rhabdomyosarcoma  Glioblastoma multiforme. Strode Note: Too many words and no image.

5. Question Does Mer play a role in the development of Acute Lymphoblastic Leukemia? Does Mer play a role in the development of Acute Lymphoblastic Leukemia? Strode Note: Avoid yes/no research questions.

6. Hypothesis I hypothesize that Mer will be abnormally expressed in the T-cell and B-cell Acute Lymphoblastic Leukemia cell lines REH and Jurkat, and certain patient samples. Strode Note: What’s wrong with this?

7. Methods Cell Culture Cell Culture is a sterile procedure to grow cells for use in different types of experiments including PCR. Cell Culture is a sterile procedure to grow cells for use in different types of experiments including PCR. Strode Note: I have no idea what “cell culture” is from this slide.

8. Methods Isolation of RNA and Preparation of cDNA In order to perform PCR, the RNA (a single stranded genetic sequence) must be extracted from the cells then turned into cDNA. This is done by exploding the cells to release its contents, digesting the DNA in the cells, then adding random decamers, which attach to pieces of RNA strands to form cDNA. In order to perform PCR, the RNA (a single stranded genetic sequence) must be extracted from the cells then turned into cDNA. This is done by exploding the cells to release its contents, digesting the DNA in the cells, then adding random decamers, which attach to pieces of RNA strands to form cDNA. Strode Note: Too many words and no image.

9. Methods Polymerase Chain Reaction (PCR) PCR is used to amplify DNA and to test for the presence of certain genes. PCR is used to amplify DNA and to test for the presence of certain genes. Two PCR primers (short pieces of single stranded DNA that match the sequences of the genes you are testing for) and Taq (an enzyme that manufactures the DNA copies) are added to the cDNA. Two PCR primers (short pieces of single stranded DNA that match the sequences of the genes you are testing for) and Taq (an enzyme that manufactures the DNA copies) are added to the cDNA. cDNA placed in a PCR machine to replicate. cDNA placed in a PCR machine to replicate. Double stranded cDNA separates, primers attach to the strands, and Taq polymerase elongates the new DNA strands. Double stranded cDNA separates, primers attach to the strands, and Taq polymerase elongates the new DNA strands. New DNA strands separate and the cycle repeats. New DNA strands separate and the cycle repeats. Amplified products run on a gel and imaged on a gel doc. Amplified products run on a gel and imaged on a gel doc. Strode Note: Too many words and no image. See the next slide.

10. Methods Strode Note: How about just this image and then walk us through how PCR works.

11. Acknowledgements Dr. Kelly Sawczyn Dr. Kelly Sawczyn Dr. Amy Keating Dr. Amy Keating Dr. Dana Salzberg Dr. Dana Salzberg Dr. Doug Graham Dr. Doug Graham University of Colorado Health Sciences Center University of Colorado Health Sciences Center Strode Note: I like putting acknowledgments first, right after the title slide.