Why no gene deletion in S. cerevisiae using Lox p system?

More Thiyagarajan Sekar's questions See All

How can I interpret this FT-IR spectra?

Hello everyone! I made a solid dispersion of valsartan using PEG 6000 and Kollidon VA64 as polymers, prepared by solvent evaporation method. For characterization, I checked my sample using FTIR...

11 May 2024 4,598 4 View

What is wrong with my DSC peak?

Hello guys, i would like to ask about my DSC thermogram results. There is an unexpected crowded peak that appeared on my thermogram result (the one in the blue circle). I have tried to search...

11 May 2024 8,480 4 View

Why I am not getting accumulated GFP in vacuole for om45 WT GFP yeast strains?

I am following the standard method which is mentioned in Tomotake Kanki et al., 2009 research paper Monitoring mitophagy in yeast: The Om45-GFP processing assay . I am growing the cells in SD-HIS...

04 May 2024 6,463 0 View

Why is my DSC thermogram crowded?

I made some solid dispersion samples with VAL (valsartan) as the active compound. I was expecting it to become amorphous form. However, when I first run it through DSC, the pure VAL drug has some...

25 April 2024 2,818 4 View

`Does freeze thaw effect will have an impact on the peptide's stability?

Hello! I am working on investigating the effect of freeze and thaw on the peptides stability. The peptide is relatively short less than 50 AA so my question is would this have an impact at all? Is...

07 April 2024 7,521 4 View

Why do I observe multiple non-specific bands and the disappearance of the protein marker in my yeast protein Western blot?

I am relatively new to conducting Western blots. I employed the alkaline lysis method to extract whole proteins from yeast. Subsequently, I introduced the Opti Protein Marker (ABM, CAT log NO:...

05 December 2023 1,418 5 View

Hi I am using a cadence virtuoso to design a two stage CMOS OP-Amp. Facing issues with dc analysis in the hspiceD simulation?

I will paste the output log file and the netlist so that if anyone could point me the fix for the issue would be really helpful. ****Output Log*** Using:...

15 April 2023 4,437 2 View

How to measure the auxin gibillin level of growth hormone in plant as actinomyces strain ?

I want to know about how to measure the growth hormone in plant in seed germination

28 February 2023 240 2 View

How to prepare sample for analysing Glass Transistion Temperature (Tg) of the Latex Polymer?

I have used wet samples and dried film samples for Tg in DSC but I could not get any major peak.

04 August 2022 8,714 2 View

How to obtain the color contours in Ansys CHEMKIN ?

After the analysis of hydrogen-air combustion i can able to obtain the plots but when am trying to take contours, i cant find any data there. Is there any method to obtain them. Do i need to...

14 February 2022 4,592 0 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 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

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,418 1 View

Are these cassettes suitable for expressing PETase mutant in E. coli?

I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...

28 July 2024 7,559 1 View

Optimal siRNA Volume for Complexation with Liposomes in a 2.5 mL Solution with 10 mg/mL Lipid Concentration?

How much volume of siRNA can I add to 2.5 mL of liposomes with a total lipid concentration of 10 mg/mL?

27 July 2024 6,920 1 View

If my gene of interest has high GC content can it be problematic in sequencing? What kind of error is expected with GC rich gene sequences??

Gene sequencing related trouble shooting

25 July 2024 4,149 2 View

Why my negative control siRNA is decreasing the target gene's expression?

Hi Everyone, I'm using an siRNA kit to knock down a target gene. The kit guarantees that the negative control doesn't target any sequence in mouse genome, and when I use BLAST I don't find any...

23 July 2024 2,673 6 View

The reason for breeding Loxp mice with FLPe mice?

I am planning to use CD4 Cre mice crossed with Loxp mice. However, the laboratory from which I am trying to get the loxp sperm says they have "the product of the EMMA ko crossed with the FLPe to...

16 July 2024 7,475 1 View

Analysis of MHC-I and II alleles with CNVs and unassigned loci?

I am working on a dataset of MHC-I and II alleles from a bird species sequenced with Illumina. We were not able to assign alleles to loci through MHC-typer as we were over the limit of 150 alleles...

15 July 2024 182 1 View

Chanuka Kulatunga

What is the promoter for Cre in your vector ?

Thiyagarajan Sekar

GAL1 promoter and I have used different Galactose concentration from 0.5 to 4 %.

Alejandro De Las Peñas

I am not famliliar with the LoxP system, but PCR a marker with Fw and Rv primers complimentary to the gene to be knockout, works out pretty well.