[DIYbio] Re: query from PBS NewsHour

On Thursday, August 30, 2012 3:41:27 PM UTC-4, Tom Randall wrote:

On Sunday, April 1, 2012 1:30:11 PM UTC-4, Tom Randall wrote:

(1) Is anyone actually doing DIY work on synthetic biology?  What makes it DIY (e.g. taking place in a community lab)?  What makes it synbio (and not just biology or molecular biology)?

Thanks,

Jason

I have a lab at home and will likely soon be doing something that loosely might be considered synbio, but only because I intend on ordering a sequence from one of the companies like Blue Heron.
Specifically the project is that I have a morphological mutant in my organism of interest, an ascomycete fungus Neurospora crassa. I am interested in determining the gene responsible for the phenotype of this mutation. It has been genetically mapped so I know within 20-40 kb where it is. I have in the past done Sanger sequencing on a candidate gene, which found no mutation in or around that particular gene so decided to simply get the whole genome sequenced. Hopefully then I can compare that sequence to reference and find a SNP/indel responsible. If that is possible, I intend to order a sequence of ~ 1 kb around this mutation, including the candidate mutant allele, and transform this into a strain with the wild type allele of this candidate gene in order to generate a replacement of the wt allele with the mutant allele by homologous recombination in order to re-create the original phenotype and thus confirm (or not) the genetic lesion that resulted in the original mutant allele. I have sent the DNA from this mutant strain to Operon, but only last week so the sequencing is still is in progress. I would say this is only loosely synbio because I am simply going to be using a synthetic construct as a convenient  tool in the context of what is really a molecular genetics/bioinformatics project.

An update on my sequencing project, has turned out nicely so far.

http://www.roningenetics.org/Sequencing.html