Should I add a promoter upstream of my CRISPR-inserted gene?

Hello, I want to insert a gene into my cells using sgRNA, Cas9, and an HDR template. The gene that I will insert into has its own promoter. Because I will cut the gene downstream of its start...

14 January 2021 1,170 2 View

What are the insilico tools available to recognize transcription binding sites on a promoter ?

I have to control the expression of a gene ( in vitro ). For designing an optimal promoter assay, I would like to know more about Transcription factor binding region, enhancer and repressor...

11 January 2021 7,992 3 View

Why negative controls also showing the specific bands of promotor and other regions (GUS, GFP etc.)?

Dear Researchers, I did a transformation of an empty vector (pCAMBIA1301/1302) into my passion fruit plants, for the optimization of the transformation protocol. After transformation; DNA...

06 January 2021 8,465 99 View

Output of rankMANOVA in R?

Dear all, I am working on data including two responses (count data, relatively correlated~R2=0.4, non-normal distribution, and violated the homogeneity of covariance) and three independent...

26 November 2020 711 3 View

Luciferase assay for gene-miRNA interaction: which vectors should be used?

23 November 2020 2,813 2 View

Inconsistent results after plasmid re-purification from frozen stock?

Hello everyone, I am performing reporter gene assays on some uORF mutants vs my WT 5' UTR of my gene of interest. I am doing double transfection on HEK293T cells, using Nanoluc as a test reporter...

22 November 2020 2,746 5 View

Luciferase assay for miRNA-gene: How to construct the mutated prime?

I have experimental evidence (qRT-PCR and WB) to suggest a miRNA downregulates a gene. To test if the regulation is direct I want to perform a luciferase reporter assay. So I designed a primer...

15 October 2020 7,641 4 View

RS dataset of ice cover of Arctic Antarctic?

I am looking for datasets of ice cover of Arctic Antarctic. I want only India related dataset. If you have any source of dataset please share with me. It will be very helpful for my...

13 October 2020 7,137 1 View

What makes two promoters compatible in a single plasmid?

Hello all. I am attempting to generate a bacterial strain that expresses two separate fluorescent proteins by placing the proteins into a single plasmid. To do so I plan on cloning the one...

16 September 2020 2,194 3 View

How much plasmid DNA should I use in my transfection?

Hi, I am trying to perform a luciferase assay using Applied Biological Materials (abm) plasmids, but I am uncertain on how much plasmid DNA should I use. I already decided that I will plate the...

11 May 2020 7,901 7 View