MLPA Automation Gemma Dennis GT Training day

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MLPA Automation Gemma DennisGT Training day
Newcastle23/11/11
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MLPA _at_ Bristol

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

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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.

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MLPA Automation

• Centralised booking system
• All requests on one sheet 96 well plate format
• All information needed for set up

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Sample No MLPA Name PCR No DNA Conc (reaction) DNA Dilution Volume DNA Volume (dilution) Water Volume (dilution) DNA Volume (reaction) Probe Mix Volume Ligase Mix Volume Ligation vol to PCR PCR Mix Volume
M.11.05592 p300ARS 153 50 20 1.82 18.18 5 1.5 16 5 20
M.11.05708 p300ARS 153 50 20 4.35 15.65 5 1.5 16 5 20
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Sample No MLPA Name PCR No DNA Conc (reaction) DNA Dilution Volume DNA Volume (dilution) Water Volume (dilution) DNA Volume (reaction) Probe Mix Volume Ligase Mix Volume Ligation vol to PCR PCR Mix Volume
M.11.05592 p300ARS 153 50 20 1.82 18.18 5 1.5 16 5 20
M.11.05708 p300ARS 153 50 20 4.35 15.65 5 1.5 16 5 20
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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

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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

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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

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• Any Questions?
• Acknowledgments
• Kenneth Smith
• Mark Greenslade
• Laura Yarram