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BMC Molecular Biology

, 11:94

First Online: 07 December 2010Received: 09 August 2010Accepted: 07 December 2010

Abstract

BackgroundAmplification of single-stranded DNA circles has wide utility for a variety of applications. The two-primer ramified rolling circle amplification RAM reaction provides exponential DNA amplification under isothermal conditions, creating a regular laddered series of double-stranded DNA products. However, the molecular mechanism of the RAM reaction remains unexplained.

ResultsA RAM reaction model predicts exponential accumulation of a double-stranded DNA product size series, and product-size ratios, that are consistent with observed RAM reaction products. The mechanism involves generation of a series of increasing size intermediate templates; those templates produce RAM products and recursively generate smaller intermediate templates. The model allows prediction of the number of rounds of circular template replication. Real-time RAM reaction data are consistent with the model. Analysis of RAM reaction products shows exponential growth limitation consistent with the model-s predictions.

ConclusionsThe model provides a rationale for the observed products of the RAM reaction, and the molecular yield among those products. Experimental results are consistent with the model.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2199-11-94 contains supplementary material, which is available to authorized users.

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Author: Thomas P Beals - James H Smith - Raymond M Nietupski - David J Lane

Source: https://link.springer.com/



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