Tag Info

I went to the Yeastbank website at Weihenstephan for some info. The keyword here is "Stamm," which is German for stem, clade, clan, or strain. So, I would take this to mean that the 34/70 is an isolate (#70) of strain 34. Two of 34/70's strengths, according to the link above are it makes clean beer and gives a pleasant taste profile due to its low yeast-like ...

A dependable protocol for yeast DNA extraction I used to use was broadly similar to the protocols you cite but included an ethanol precipitation before and after the P:C extractions. The only expensive material was time. The general outline was: Alkaline lysis Ethanol precipitation RNase A treatment of resuspended DNA for 15 minutes Phenol:chloroform ...

I am not sure why you say there is no information... a quick Google search returned a few interesting pages... In this paper: Progress in Metabolic Engineering of Saccharomyces cerevisiae - Nevoigt, Microbiol Mol Biol Rev. 2008 the author says: The identification of the entire genomic sequence of a commonly used lager brewer's yeast strain, i.e., ...

I'd take that computational angle and run the sequence data through tRNAscan-SE (Lowe & Eddy, Nucl Acids Res 25: 955-964). Ideally, you'd install this locally. This tool is what the UCSC folks use and it has been the best known, most widely used tRNA predictor for years. It's what we all used on Arabidopsis thaliana genome annotation back in the late ...

It depends upon exactly what you have done. The standard way of using a yeast intergrating plasmid (YIp) is for it to integrate into the genome by recombination between a piece of yeast DNA that the YIp carries and the same DNA in the genome. This is often, but not necessarily, the selectable marker. So for example a YIp carrying the URA3 gene as its only ...

Yeast integrating plasmids are known to be stable, even in absence of a selective medium but can revert like most homologous recombination plasmids. Quoting from the book Yeast Gene Analysis (pg476): YIps are generally more stable than YRp or YEp plasmids. As a result it is safe to grow most YIp-bearing strains in rich media, although it is good ...

I think that you could try a similar approach to GSFS: use transduction in proteins (if you don't know star code, then you must use 3 strings for each gene) use a basic tool (a stand alone like UNIPROT tools) to identify the proteic domain type (chain alpha, ..) divide the genes by proteic domain type (pdt): which contains which pdt and the pdt order ...

http://www.protocol-online.org/biology-forums/posts/26634.html I found this link - it says 'we don't really know'. It says PEG binds DNA, I assume shielding the membrane from its negative charge and allowing internalization to happen. I would guess that the amphipathic nature of PEG, being partly hydrophobic, also helps soften up the membrane. ...

I would use an RNA microarray to look at those difference instead of sequencing. To delicately amplify your tRNAs in an unbiased manner would be a tricky molecular biology endeavor. I wouldn't be surprised if there were detectable and significant differences. Those experiments will be able to confirm your hypotheses regarding codon usage bias.

I've successfully used the Zymoprep kit, but only for smaller plasmids in 2-hybrid experiments. So their cell lysis enzyme/buffer system works well, but I'd guess that your 42kb plasmid is precipitating out with the genomic DNA. The standard Qiagen midi columns are capable of capturing large constructs. This has worked wonders for me when I was purifying ...

According to this site, for yeast (I assume this means Saccharomyces cerevisiae) you should use 50 µg/ml to 200 µg/ml; for fungi (!) use 100 µg/ml to 300 µg/ml. The site also stresses the importance of the pH of the medium. I think that, unless a Hansenula expert comes along, you will have to try an initial experiment to measure the sensitivity of ...