PRE-CONGRESS WORKSHOP:
HANDS-ON WORKSHOP ON PREIMPLANTATION HLA TYPING
Reproductive Genetic Institute (RGI)
Chicago, Illinois, U.S.A.
Register for Pre-Congress workshop: Preimplantation HLA Typing. |
Dates: April 19 - 22, 2009
Cost: $2,300
Faculty:
Joe Leigh Simpson, MD, RGI Medical Director, President of PGDIS
Yury Verlinsky, PhD, Director of RGI
Svetlana Rechitsky, PhD, Director of RGI DNA Laboratory
Oleg Verlinsky, MS, Deputy Director of RGI DNA Laboratory
Tatyana Sharapova, BS, RGI DNA Laboratory
Anver Kuliev, MD, PhD, Executive Director of PGDIS
Sunday, April, 19: Arrival and Registration
Monday, April 20: PGD strategy for HLA genotyping and mutation analysis
| 9:00 - 9:15 |
Opening of Workshop
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| 9:15 - 10:15 |
Introductory lecture
Basic Principals of Single Cell Analysis involving Preimplantation HLA Typing: Problems and Solutions
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| 10:15 - 10:30 |
Coffee Break and Discussion |
| 10:30 - 12:00 |
PGD Molecular Laboratory set up
Equipment and chemicals required for Preimplantation HLA Typing
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12:00 - 12:30
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Instruction for the start of Hands-On Procedures |
12:30 - 14:00
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LUNCH & DISCUSSION |
| 14:00 - 16:00 |
Amplification of HLA markers, mutant and normal alleles from single cells
All participants will be divided in groups of two and start working on single cell amplification
Review of PCR protocol for the first round amplification
Start single cell lysis procedure and perform Hot Start. First round (takes about 3, 5 hours)
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| 16:00 - 18:00 |
Haplotype assignment for a couple requesting HLA genotyping and mutation analysis
Raw Data review and analysis – a table and pedigree preparation:
Each participant will be provided with fluorescent STR printouts and mutation analysis, to be able to create a table and family pedigree, and to establish a strategy for the particular PGD case, under supervision of moderator |
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Tuesday, April 21 Preimplantation HLA Typing with and without PGD
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| 9:00 - 10:00 |
State of the Art Lecture: Present Experience in Preimplantation HLA Testing
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| 10:00 - 12:30 |
Proceed with single cell amplification
Review PCR protocol for the second round of amplification
Prepare titer plates with PCR buffer for the second round of amplification
Transfer PCR product from the first round tubes into the titer plates
Start second round of amplification
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12:30 - 14:00
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LUNCH & DISCUSSION |
| 14:00 - 18:00 |
Run PAAG electrophoresis for evaluation of amplification efficiency
Perform Restriction digestion of PCR product for the mutation analysis
Start sample preparation for fluorescent genotyping and run fluorescent electrophoresis
Run PAAG electrophoresis for evaluation of amplification efficiency
Print STR results |
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Wednesday, April 22: Data Review, Table Preparation, Genotypes Assignment and Recommendations for the Physician
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9:00 - 12:00
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Each participant performs the demonstrated techniques under supervision of the moderator |
12:00 - 13:00
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General Discussion and Closing Remarks |
Register for Pre-Congress workshop: Preimplantation HLA Typing at RGI
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