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 8,865 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 10,031 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 2,572 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 3,474 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 4,366 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 2,793 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 5,431 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 3,734 7 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 8,691 31 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 5,982 9 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you
08 August 2024 4,733 2 View
I have prepared a liquid broth containing Yeast (S. Cerevisiae) that i need to add from it to fermentation media of lignocellulosic hydrolysate. I need to know based on what parameters do we add...
06 August 2024 662 1 View
Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...
06 August 2024 1,495 2 View
After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...
05 August 2024 9,637 2 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
Brain and body mass together are positively correlated with lifespan (Hofman 1993). The duration of neural development is one of the best predictors of brain size, and conception is the best...
05 August 2024 6,247 3 View
kindly reply me. Thanking you in advance.
05 August 2024 7,727 4 View
Is it possible to conduct a molecular dynamics simulation to see the effects of a specific carbohydrate on the structure of lipids (e.g., micelle structure)? I am a beginner in this field and plan...
03 August 2024 3,371 3 View