Do you know a protocol for chemical transformation for Debaryomyces hansenii?

02 February 2014 1 3K Report

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. Can anyone suggest a relatively efficient protocol for chemical transformation of yeast D. hansenii?

Badges
Science topic

More Dmitry S Karpov's questions See All

Do you know efficient DNA sequences for plasmid nuclear import and how they work?

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

Are there simple and efficient methods for extraction both RNA and DNA from the same sample of Gram-positive bacteria?

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

Can you suggest software or on-line server to annotate Drosophila genome mutations?

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

What do you do if you have troubles with cloning into a plasmid vector?

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

What could be the reason behind an unexpectedly low weight band of 3xHA tagged large protein on western-blot?

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

Is it possible for a mutation in the yeast genome to be repaired upon URA3 marker excision?

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

How to optimize conditions for expression of chimeric membrane proteins in E.coli?

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

Which media is the best for high level protein production in E.coli?

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

Can LB media inhibit IPTG-induced protein expression?

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

What is the efficient method for blocking total protein translation in yeasts?

We tried to make cycloheximide chase assay in yeasts and found that even 200 mkg/ml concentration of cycloheximide in complete selective media does not stop our protein translation which is very...

11 December 2012 1,877 5 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

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

fatal error

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

fatal error

16 June 2021 0 0 View

What is the most Operating System (OS) which can be INSTALL on Tiny hard devices like Raspberry Pi (RPi) hardware ?

I'm targeting to deploy a mesh network and manually configure MANET routing protocols. I'm preparing scenarios, architectures, and hard devices needed to do that. Are there some step-by-step...

03 March 2021 1,931 5 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

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

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 View

How to repeat an operation on an atom selection for multiple times in PyMol?

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