2D-PCR: a method of mapping DNA in tissue sections

Michael Armani; Jaime Rodriguez-Canales; John Gillespie; Michael Tangrea; Heidi Erickson; Michael R. Emmert-Buck; Benjamin Shapiro; Elisabeth Smela

doi:10.1039/B910807F

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Lab on a Chip

Issue 24, 2009

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2D-PCR: a method of mapping DNA in tissue sections

Michael Armani , Jaime Rodriguez-Canales , John Gillespie , Michael Tangrea , Heidi Erickson , Michael R. Emmert-Buck , Benjamin Shapiro and Elisabeth Smela

Lab Chip, 2009, 9, 3526-3534

DOI: 10.1039/B910807F
Received 02 Jun 2009, Accepted 20 Aug 2009
First published on the web 12 Oct 2009

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A novel approach was developed for mapping the location of target DNA in tissue sections. The method combines a high-density, multi-well plate with an innovative single-tube procedure to directly extract, amplify, and detect the DNA in parallel while maintaining the two-dimensional (2D) architecture of the tissue. A 2D map of the gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was created from a tissue section and shown to correlate with the spatial area of the sample. It is anticipated that this approach may be easily adapted to assess the status of multiple genes within tissue sections, yielding a molecular map that directly correlates with the histology of the sample. This will provide investigators with a new tool to interrogate the molecular heterogeneity of tissue specimens.

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Supplementary Info

A figure showing the pressure compression rig, theelectrophoresis gel validations of the 2D-PCR method, and completedetails of the miniaturization studies, including themicrofabrication, reagent loading by capillary action, cross-talkstudies, demonstration of PCR, and discussion
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Michael Armani
Jaime Rodriguez-Canales
John Gillespie
Michael Tangrea
Heidi Erickson
Michael R. Emmert-Buck
Benjamin Shapiro
Elisabeth Smela

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