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DNA PREPARATION FOR THE ISRAELI BONE MARROW REGISTRY: SUCCESSFUL INTEGRATION OF THE ABBOTT M1000 MOLECULAR WORKSTATION AND THE TEPNEL/LIFECODES MULTIPLEX PCR KIT FOR HIGH THROUGHPUT MOLECULAR HLA TYPING.
Ariane Davidson PhD 1, Bracha Shainberg PhD 1, George Prajgrod PhD 1, Dina Noff PhD 1, Dafna Lipinsky PhD 2 and Tirza Klein PhD 1. 1 Tissue Typing Unit, Central Lab. Maccabi Health Care Services, Rechovot, Israel and 2 Abbot-Ilex Medical .
Since BM transplantation began placing an increasing demand on donor resources in Israel, drives were initiated 5 years ago that provided >200,000 BMDW registered donors. The registry is served by a new laboratory, of the Maccabi Health Care, capable of tissue typing with high throughput.
HLA typing requires a large yield of highly purified genomic DNA, required for PCR-SSOP and SSP typing, and archiving DNA as basis for on-demand high resolution typing. We report our initial experiences using the Abbott m1000 workstation for DNA extraction from our last 14,000 drive. In addition, this is the first documented integration of the workstation with the Tepnel/Lifecodes platform for low resolution typing. This fully automated system easily extracted DNA from 144 
primary tubes
per work-day, i.e. 48 x 3 cycles, but can potentially process 288 specimens/day. For 400
l of whole blood, the median yield of recovered DNA is 14 g in a final volume of 400l, giving a DNA concentration of 30 ng/l. The DNA was typed for low resolution A*B*DRB1* using the Tepnel/Lifecodes Multiplex PCR kit with optimized protocols. High resolution typing utilized the One Lambda PCR-SSP kits.
Conclusions: The Abbott m1000 DNA extraction workstation is a valuable and reliable system for high throughput DNA preparation. Once the operational parameters have been optimized, and strictly adhered to, it can be utilized for most HLA laboratory applications and registry DNA archiving. Additionally, Tepnel/Lifecodes Multiplex platform can be easily integrated with the workstation
s operation.