Please, suggest a method for estimation (quantification) of DNA repair effeciency (general or specific pathways) in yeasts Saccharomyces cerevisiae.
Non homologous end joining essays using tissue extracts is performed as per the attached paper. You might be able to follow the same with yeast extract as well.
As I know there are particular DNA sequences that mediate efficient import of plasmids in into the nucleus, for example, cPPT sequence of the lentiviral vectors (one of its functions is...
05 June 2018 8,741 2 View
I am working with highly mutagenized Bacilli species and wish to measure expression level of my favorite genes by real-time PCR. As I know, there are no reliable internal reference genes for...
31 December 2017 9,894 1 View
I have a tab-delimited list with genomic mutations of Drosophila melanogaster mapped to FlyBase r5.50 genome assembly. List contains such fields as gene name, chromosome, strand, genomic...
03 April 2015 2,456 5 View
I am trying to delete a gene of interest in D.hansenii. There is a protocol for transformation of this yeast by electroporation, but for me it is more convenient to use chemical transformation....
01 February 2014 3,360 1 View
My colleagues and I have an experience of successful cloning into various plasmid vectors. We noticed that it is hard to clone some inserts into one vectors, while the same inserts can be readily...
09 October 2013 4,233 6 View
We have cloned a yeast gene for a large protein (ca mass about 93 kDa) together with its native promoter into plasmid where our protein is C-terminally tagged with 3xHA epitope. Insert together...
09 October 2013 2,664 17 View
In yeast genome we have introduced a mutation which replaces transcription factor (TF) binding site to site for restriction endonuclease. The integration marker is URA3. The correctness of marker...
09 October 2013 5,304 6 View
I try to display an enzyme on the surface of E.coli cells. To do this I have made a chimeric protein consisting of signal peptide (SP) - enzyme (E) - outer membrane protein of E.coli (MP). The...
08 September 2013 3,609 7 View
Standard complex media like LB that is routinely used for cultivation of E.coli and performing various experiments including protein expression has several serious limitations. For example,...
08 September 2013 5,829 9 View
I failed to induce expression of my protein in BL21(pLysS) from pET11c. I obtain the cell culture by growing individual colonies from the plate on LB to OD600 0.5-1. Protein expression is induced...
08 September 2013 8,520 31 View
How to calculate the RMSD values for a MD simulation using MOE?
07 August 2021 0 0 View
When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.
16 June 2021 0 0 View
I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance
03 March 2021 3,568 1 View
Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...
03 March 2021 5,978 5 View
Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...
02 March 2021 4,569 4 View
Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...
02 March 2021 3,024 2 View