1 / 10

MLPA Automation Gemma Dennis GT Training day – Newcastle 23/11/11

MLPA Automation Gemma Dennis GT Training day – Newcastle 23/11/11. MLPA @ Bristol. CMT (Charcot-Marie Tooth) Telomeres FH (Familial Hypercholesterolaemia) SMA (Spinal Muscular Atrophy) ARS (Axenfeld-Rieger Syndrome) TAR (Thrombocytopenia-Absent Radius syndrome). Why Automate?.

turi
Download Presentation

MLPA Automation Gemma Dennis GT Training day – Newcastle 23/11/11

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. MLPA AutomationGemma DennisGT Training day – Newcastle23/11/11

  2. MLPA @ Bristol • CMT (Charcot-Marie Tooth) • Telomeres • FH (Familial Hypercholesterolaemia) • SMA (Spinal Muscular Atrophy) • ARS (Axenfeld-Rieger Syndrome) • TAR (Thrombocytopenia-Absent Radius syndrome)

  3. Why Automate? • Solid Tissue screening by MLPA was introduced in July 2010, resulting in a large increase in sample numbers • Set-ups became complex and time consuming • Result quality was reduced due to the large run size • Automation = increased samples per set up, less staff time = cost saving.

  4. MLPA Automation • Centralised booking system • All requests on one sheet – 96 well plate format • All information needed for set up

  5. 4. Denature DNA on Thermocycler DNA Rack 1 (4 x 6) 3. Transfer normalised DNA (5ul) 2. Transfer DNA (1ul >) 5. Transfer Probemix (1.5-3ul) 6. Overnight hybridisation on thermocycler 1. Transfer Water (1-29ul) DNA Dilution Plate Hybridisation Plate Thermocycler Water Reservoir Probe Mastermixes

  6. 9. Transfer Ligated MLPA Product (5ul) 7. Transfer Ligase Mix (16-32ul) 8. Ligation Rx on Thermocycler Reagent Rack PCR Plate Hybridisation Plate 8. Transfer PCR Mastermix (20ul) 10. PCR Rx on Thermocycler Thermocycler Thermocycler 11. Load on Analyser

  7. MLPA Automation – Validation work • Initial development work based on existing Biomek workflows • Liquid handling was tested using coloured dyes (volume range 1.5-32µl) • Single kit was run to asses if adding probe mix, ligase mix and/or PCR mix at room temperature had any adverse affects • Large scale test was set up simultaneously with manual set up

  8. MLPA Automation – Observed results • Robot was able to pipette all volume sizes accurately • There were no adverse affects to all mixes being added at room temperature • All results from automated set up were comparable to those from manual set up

  9. MLPA Automation - Conclusions • Automation allows multiple MLPA tests to be set up simultaneously using a single plate • Technical work is simplified and can be set up in less time • System has ability for further tests to be added to the workflow without additional staff time being required

  10. Any Questions? Acknowledgments Kenneth Smith Mark Greenslade Laura Yarram

More Related