Is it possible to transfer a gene flanked between attL1 & attL2 of Entry vector to destination vector with attR2 & attR4, using clonase reaction ?

More Alaa Refaat's questions See All

Could anyone recommend software for drawing the physical structure of nanoparticles?

I am looking for professional software to draw the physical structure of nanoparticles or vesicles, not the chemical structures for which we use ChemDraw, but the actual physical structures,...

23 June 2024 1,292 3 View

How could I analyze the results of caspase-glo 3/7 assay?

I have done caspase-glo 3/7 assay which measures caspase activity after cleavage and generation of luminescent signal produced by luciferase. However,after I got the results, I am not sure how to...

02 June 2024 7,833 2 View

How can I identify an unknown gene of known activity ?

Hi, I've observed a new biological activity in certain cells, likely induced by an enzyme. How can I determine the gene or genes responsible for expressing this unknown enzyme? Thanks in advance

31 May 2024 9,111 3 View

Hello everyone, I am looking for collaboration to do clinical research?

I have many skills like data analysis with R programming or Spss , scientific writing and paper publishing

23 May 2024 1,683 3 View

Does GFP interferes with absorbance in MTT assay or luminescence in Caspase 3/7 Glo assay?

I have cells that produce GFP as a control, and I intend to perform the MTT and Caspase 3/7 Glo assays. The emission wavelength of GFP is 508 nm, and its absorption is 475 nm. In addition, I will...

16 April 2024 3,537 2 View

I would like to know why I obtained such false positive curves, which appears on both linear ang log scales healthy curves?

The positive curves for E coli different strains from mucual stool sample.

29 March 2024 5,907 0 View

Methylation of spata 6 gene in rat males?

methylation of spata 6 gene and its role on male infertility of rat

08 March 2024 9,667 0 View

How to prepare concentration MPs for field scale ( phytoremediation of floating plants ( water lily, water mimosa, water hyacinth )?

how to prepare concentration MPs for field scale ( phytoremediation of floating plants ( water lily, water mimosa, water hyacinth )?

04 March 2024 1,822 0 View

How to choose the suitable labtop?

I am a researcher who decided to buy labtop suitable for my work as a simulation and data analysis work Please suggest one

23 February 2024 3,635 2 View

What is the maximum duration for storage of isolated extracellular vesicles from serum in -80 for the purpose of protein content assessment?

Note: Serum isolated extracellular vesicles using immunoprecipitation (magnetic beads)

20 February 2024 4,894 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

How can we now if the promotor in the vector is well?

Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...

02 August 2024 3,987 1 View

Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View

Pink bacterial colonies?

I am cloning an overexpression plasmid with my protein of interest tagged with mScarlet. After transforming my ligated product into DH5α bacteria and plating on LB agar, I noticed colonies with a...

22 July 2024 5,953 6 View

What are the strategies to Enhance IgG-Producing Plasma Cells in mice for Monoclonal Antibody Development?

I have immunized BalB/C mice with a protein using the intradermal (ID) method with Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA), following a 14-day interval and three...

22 July 2024 9,160 2 View

The best source for amplification of ADAM17 prodomain?

hi every one I am making vector construction (for fusion proteins) and in this moment I wanna to amplification of ADAM17 prodomain with PCR. to yet, I couldn't amplified the ADAM17 prodomain with...

21 July 2024 8,660 1 View

Can I detect the FLAG sequence of my AAV vector by qPCR?

I am treating human cells with an AAV vector encoding for my gene of interest tagged by FLAG. I want to quantify the mRNA expression of my gene of interest via qPCR to discriminate the endogenous...

14 July 2024 8,482 3 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

Stable cell line generation: GOI is detected by qPCR but protein is not expressed?

Hello, I'm generating stable cell lines by transfecting plasmid DNA with lipofectamine 3000. The cells went through antibiotics selection for 3 weeks (polyclonal selection) and when I analyzed...

11 July 2024 4,812 5 View

I have constucted a plasmid vector from puc19 with no antibiotic and lacz gene. The problem is screening for transformed cells. Any suggestions?

Get too many colonies, which are mostly non-transformants when tested by colony PCR

11 July 2024 6,775 3 View