De novo DNA Synthesis using Single Molecule PCR (BioSysBio 2009)

T Ben Yehezkel et al.
Weizmann Institute of Science

When looking at the number of clones needed to sequenced in order to get one error-free molecule, the proportion of perfect molecules decrease exponentially with length. They have an error-correction system that has drastically improved this situation. They don't look for an error-free clone: they look at all of them, and the error-free ones are dispersed randomly among the clones. They PCR'ed out the error-free parts - they get an error-free sequence from looking at low-error clones. But still, cloning is a major bottleneck. So, how will in vitro clonal amplification make lives easier? In contrast with in vivo, it scales well, it automates well, it is 3-4 hours rather than 1-2 days, and it costs much less.

smPCR can integrate into recursive and other construction technologies. However, there are a few challenges. For instance, primer selection is crucial in smPCR. They construct the DNA completely automatically.

Personal Comment: The video of the automatic dna construction was a great addition to the talk.

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Please note that this post is merely my notes on the presentation. They are not guaranteed to be correct, and unless explicitly stated are not my opinions. They do not reflect the opinions of my employers. Any errors you can happily assume to be mine and no-one else's. I'm happy to correct any errors you may spot - just let me know!