How vitamin D which is unstable can be stored in 37 c incubator after treating cell line?

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Why the clear zone indicating amylase activity on agar plates dissappears after some hours?

I did qualitative assay on agar plates with starch as sole carbon source for checking amylase activity, once I added iodine the clear zone appeared arround the colonies but after some hours and...

13 May 2024 6,529 2 View

My recombinant enzyme activity is acting weird, does anyone have an explanation, or even know how to solve this problem?

I'm working with recombinant protein, it's a protease enzyme that we use E. coli to synthesize, and usually, it's stable for up to six months in the -80 freezer. Recently, the protein activity has...

17 April 2024 4,262 3 View

Sub-cloning not working?

I am a graduate student doing sub cloning of two similar genes into pBC SK(+); one of the gene was inserted but few colonies appeared on the agar after transformation, the other is not inserting...

22 February 2024 7,237 5 View

How can audiologists enhance patient-centered care in their practice?

In light of recent research findings and advancements in communication strategies for individuals with hearing loss, how can audiologists enhance patient-centered care in their practice? Are there...

17 December 2023 2,894 1 View

Why does performance point still in elastic range ?

I have a four-story steel structure When conducting a pushover analysis and examining the results, the structure's ductility ratio at the performance point did not exceed 1, and the equivalent...

08 November 2023 934 4 View

Cadmium sulfide nanoparticles from banana extract , what is the problem?

I'm preparing Cadmium sulfide nanoparticles using banana extract as a capping agent, after drying it, it looks like thin sheets, cannot be grinded as powder (The last time, it does not look like...

12 June 2023 3,604 1 View

Safe cities for women with disabilities ?

Safe cities for women with disabilities

13 May 2023 8,560 3 View

Can I use QIAamp DNA Stool Mini Kit for DNA extraction from a bacterial colony? if anyone tried this and what are the necessary protocol adjustments?

#DNA_extraction #QIAamp #Qiagen #Colony_DNA

04 May 2023 4,548 2 View

How to solve the problem if the sensor is not capable of transmitting data to base station in WSN. Is there any paper talking about this issue?

25 January 2023 5,399 4 View

Why enzymes couldnot migrate from wells to stacking gel in electrophoresis? Is there is problem in its precipitation?

i have precipitated enzyme by acetone.. when i use coomassie staining in native gel electrophoresis, it couldnot migrate to stacking and then resolving gel. is there is a problem in acetone...

10 January 2023 2,353 5 View

Unexpected 35 kDa Band Observed During Expression and Purification of Dengue 3 NS5 RdRp Domain ?

Background: I am developing an RdRp assay for Dengue virus-3 and need to express and isolate the NS5 protein's RdRp domain (~101 kDa). My expression vector includes a GST tag followed by a 6x His...

13 July 2024 9,950 3 View

What could be the expression profile of alkaline phosphatase from lactic acid bacteria?

I have already cloned ALP gene of L rhamnosus into pET22b+ vector. Cloning is already confirmed by restriction digestion and sequencing. Cloning is achieved in BL21(DE3), Codon+, Rosetta and Lemo...

02 July 2024 5,332 3 View

Can I do IPTG induction on OD600=2.6 in TB?

I have been looking at papers for protein expression and trying to follow the process. I'm using TB medium to do the process, and the paper says to grow to an OD600 = 1.8 at 37 degrees, then let...

30 June 2024 6,502 5 View

How to incease the yiald of recombinant protein expression ?

Hello esteemed scientists, I am endeavoring to increase the yield of my recombinant protein. It is a 120 kDa protein tagged with MBP and features three intrinsically disordered regions with pI...

12 June 2024 9,616 8 View

Which statistical test for mRNA stability changes upon treatments ?

Hello, I discovered that a compound that I use, which get integrated into RNA, might have an unspecific impact on mRNA stability of a gene of interest. To confirm that, I pretreated my cells for...

10 June 2024 276 2 View

Can I use richer media to increase bacterial expression of human protein?

We're using LB media, 1mM IPTG, and letting it incubate at 18°C over night. Will adding glucose or something else to the media increase bacterial expression?

06 June 2024 6,071 1 View

What guidelines, suggestions, and advice do you have for building a successful research and development (R&D) career?

Hello everyone, I am currently 26 years old (feeling like I am making a lot of mistakes at this young age), finishing my Master's degree, and planning to continue onto a PhD and possibly a...

30 May 2024 6,980 2 View

How much glucose to add to LB media to prevent leaky expression?

I am already using BL21(DE3)pLysS cells but I am still getting leaky expression in pre-expression samples before IPTG is added. LB broth is tryptone, salt and yeast. Thanks :)

28 May 2024 2,820 2 View

MCherry tag being photobleached during expression?

I am expressing an mCherry tagged protein. Yield of total protein is good but the fluorescence intensity is very low (ie. Protein assays and gels show a lot of protein but this does not correspond...

28 May 2024 4,828 1 View

How Can I Understand the Different Results from MacConkey Agar and β-Galactosidase Assays in a LexA-Based Bacterial-Two Hybrid System?

I'm conducting protein-protein interaction studies using a LexA-based Bacterial-Two Hybrid system with E. coli reporter strains. We employ MacConkey agar and β-galactosidase assays to evaluate the...

26 May 2024 4,647 0 View