I am having trouble working with G418 selection in Pichia pastoris, does any one have experience with this?

27 April 2021 1 236 Report

I have carried out concentration curves between 0,5 ug/ml and 2 ug/ml in order to determine the Minimal Inhibitory Concentration with no luck. I am working with recombinant strains. Any help is most welcome, many thanks!

Celina Galles

Thanks for your kind answer Stefan Zimmermann

! I will be checking out the paper you reccomend, but it clearly looks like we should try much higher concentrations...

Best regards!

Celina

More Celina Galles's questions See All

Why do my cells turn gelatinous upon addition of lysis buffer?

Hello, I work with HaCaT cells, and until now there hasn’t been any issues. But recently I’ve had a problem when lysing the cells. Upon the addition of lysis buffer, the cell “suspension” turns...

28 November 2023 5,248 4 View

Sac-B selection in E coli troubleshooting?

Hello, I need help improving my counterselection procedure when using sac-B. I have grown my E coli strain of interest in LB-agar media with and without sucrose (5%), at 30°C, following the...

08 November 2023 2,292 3 View

Is there a way to estimate doses for systemic/oral administration based on topical concentrations?

Hello, I am setting up a mouse study to test the effects of a number of different compounds in our mouse model. We currently have data on effective concentrations given through topical...

24 May 2021 1,706 3 View

Can someone help me get this paper?

I am doing a school project and I would really like to have this article includedReview of Microfluidic Approaches for Fabricating Intelligen... The issue is my university does not have a...

18 May 2021 3,977 5 View

In theory, can metabolic diseases, characterized by the lack of production of a protein, be treated (not cured) by supplementing with that protein?

I was reading about metabolic disorders caused by the lack of production of a protein. Can a possible treatment be to supplement the patient with that lacking protein, or would the body degrade it...

19 November 2020 902 2 View

Can you please recommend a protocol for native protein deglycosylation using PNGase F?

I was able to deglycosylate the denatured version of our protein of interest, but saw no deglycosylation when I tried with the native protein. Any tips?

10 December 2019 3,201 0 View

Can anyone help with a load-controlled triaxial test using clay plasticity material model?

I am attempting to simulate a triaxial test in ABAQUS/Standard using the clay plasticity model in the ABAQUS material database. I am using a 3-D cylindrical specimen which contains a single...

03 October 2014 6,998 3 View

What role do research and statistics play in Organizational Psychology?

I am examining the field of Organizational psychology and the difference between practicing OP's and research OP's.

01 February 2013 6,645 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

How can microbial engineering techniques be used to enhance the functionality and efficacy of probiotics ?

through genetic modification or metabolic engineering? Is it helpful, good for health ?

22 July 2024 7,811 4 View

ATG in pPICZalfphaA vector: Yes or Not in front of the gene of interest ... ?

Is it appropriate to place the ATG codon in front of the gene of interest, since there is a secretion signal that has its own ATG in front of this gene? I need my protein of interest to be...

10 July 2024 3,050 5 View

Is there any way to calculate whole genome similarity of organism without the whole genome sequencing ?

is there any ways that i can find genome similarity of an organism without whole genome sequencing ,like using maths formulas or experimental progress ?

05 July 2024 4,070 3 View

Is it okay to grow bacteria in 30 for cloning? What is the lowest mass of Vecor I can take for cloning? what should be the optimum insert ratio?

I am using a vector that has repeat sequences, and using Mach1 competent cells. I heard that if the cells are not stable cells, then we shouldn't grow them in 37 to avoid recombination to happen...

26 June 2024 3,735 3 View

How to replace Zeocin resistance gene in Pichia genome after integration?

I have been using Zeocin marker for target protein expression to verify integration in Pichia system. However, I would like to remove Zeocin marker since it's antibiotic related and not ideal for...

15 June 2024 3,851 2 View

Why is the blue and white spot screen negative for all white spots?

The protein was expressed using the insect cell system, and the recombinant plasmid was successfully constructed as verified by sequencing, and the recombinant plasmid was transformed into DH10...

13 June 2024 2,590 4 View

I want to design a study for ex situ conservation of rare ferns based on ecological and genetic diversity. what could be possible method?

I want to design a study for ex situ conservation of rare ferns based on ecological and genetic diversity. what could be possible method other than cryopreservation of spore for a short term...

10 June 2024 4,992 1 View

Is a 5` phosphate needed for homologous recombination?

Is a 5` phosphate needed for homologous recombination? I am a bit confused whether phosphorylated DNA is needed to achieve efficient homologous recombination in e coli. I believe there are methods...

06 June 2024 7,065 1 View

How does one get and add a DOI to an existing file upload?

to "Familiar biological, chemical and physical events credibly evolve the Standard Genetic Code", for example?

06 June 2024 5,095 1 View