Can I put a WPRE upstream of IRES in a bicistronic vector?

07 August 2023 0 8K Report

I am interested in creating a bicistronic vector in which the WPRE follow the first gene and is upstream of the IRES rather than following the second gene and downsteam of the IRES. Can you point to any examples where this has been implemented and did it result improved expression of the first gene.

Badges
Science topic

More Mike Mcdonald's questions See All

Who of all of you will help TPEOM to re-originate the Earths atmosphere to it original 5 atmospheric elements/gases?

Global ‘Weather’ has stopped, as ‘aqua thermionic instituted by the ‘Global thermo atmospherics has instituted global thermodynamics of 120º C + and thus all waters are, or have boiled into a...

06 August 2024 8,714 0 View

Who of all the Global Scientific community will help me Prof. Dr. Yoshida make way for TPEOM, MEC ~EMC to return the atmospheric gases to the norma ?

TEP presentation caption (The Environmental Project) Re: Why should Washington’s DC, or any country government point of location think of as nowadays of as to being 'tomorrow as to come! if it...

03 August 2024 2,483 1 View

PH is wrong making up Laemmli running buffer; why?

I found the following recipe at Abcam and converted it to 5x to make a stock solution. Running buffer (Tris-Glycine/SDS) 25 mM Tris base 190 mM glycine 0.1% SDS Check the pH and adjust to...

31 May 2024 1,444 2 View

What is your answer, decision for this?

Who of all of you that see, read this will make action' with me to save this world from its NOW happening global thermodynamics of of a very bad exterminating start of 01/05/2047, and into a...

09 March 2024 1,039 18 View

I am looking for datasets containing DXA body composition and anthropometric data in children?

I am looking for datasets to supplement the NHANES datasets.

09 February 2024 1,471 0 View

Non-radioactive EMSA with biotinylated oligo detection by strep-HRP and/or strep-alkaline phosphatase?

Looking to perform a non-radioactive EMSA to study protein binding to ssDNA. Before doing so, I’m looking to optimize detection. The two detection series are: 1) biotinylated oligo, strep-HRP...

01 February 2024 8,160 0 View

Is the researchgate member "Budee U Zaman" a scammer or a bot?

Dear colleagues and members, the researchgate member Budee U Zaman (researchgate.net) copied one of my preprints one-to-one and passed it off as his own work. He only slightly changed the title....

04 January 2024 4,312 1 View

How can I measure hesperidin content of fruit sample Citrus reticulata other than HPLC?

Good day! I am trying to find a way to measure hesperidin content of the peels of my fruit sample (Citrus reticulata), other than HPLC. I plan to ferment my peels with food-derived mixed cultures...

13 October 2023 1,248 2 View

Can you compile a calibration curve with a standard that contains various compounds?

Can you compile a calibration curve with a standard that contains various compounds? For example a fatty acid methyl ester mix given that you know the concentration of each fatty acid in the mix?...

03 October 2023 2,108 1 View

Buffer that works with Primestar GXL?

Anyone know of a buffer that will work adequately for PCR with Takara's Primestar GXL? I have tons of enzyme but am out of buffer. Takara will sell a pack of extra buffer tubes, but it won't...

12 September 2023 2,451 1 View

How to load and plot 2D-PIV (particle image velocimetry) recorded velocity vector field in Techplot?

Dear Researchers I need to know, how to load and plot 2D-PIV (particle image velocimetry) recorded velocity vector field data in Tecplot? Thank You

02 August 2024 3,615 1 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,891 4 View

I need help with cloning a 200 bp insert and a 4,7 Kb pEGFP-N1 vector. How can I do it?

I've been trying to clone my N-terminal inserts in the comercial pEGFP-N1 vector. Initially, I cloned my N-terminal insert into a pGEMT-easy vector to ensure that the insert digestion was done...

09 July 2024 4,276 1 View

Cloning restrction digestion needed before sequencing?

Hello, recently i am an issue swith cloning where I am using the subclone method to insert my gene length is 1.5kb and my vector(pMAL-c6T) is 5kb I got the colonies which i screened through the...

04 July 2024 7,977 2 View

Why I am unable to get the expected insert at the cloning site?

I am trying to clone a gRNA (23 bases) into the Crispr cas9 pDirect23A vector by using restriction enzyme cloning. I have performed a complete experiment, starting with plasmid isolation,...

02 July 2024 5,447 2 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

List Plasmid suitable for Homology cassette in CRISPR-Cas9 protocol for Knock-In in mammalian cell line ?

Hello, I am working on creating a mutant endothelial cell line having Neomycin knock-in present, I have designed my homology arms of 1kb size having neomycin in between them, I would like to...

25 June 2024 1,195 0 View

Can you provide me latest literature ?

MY RESEARCH TOPIC IS THE IMPACTS OF KIRI DAM ON SWEET POTATO CULTIVATION FOR FARMERS OF THE UPSTREAM COMMUNITIES, ADAMAWA STATE, NIGERIA. I NEED RELATED AND LATEST ARTICLES (2020 TO DATE) TO...

22 June 2024 4,116 1 View

How can I convert remote sensing (RS) images to vector data automatically?

I am working on a project that involves analyzing remote sensing (RS) images. I need to convert these images into vector data for further analysis. I am looking for automated methods or software...

22 June 2024 6,165 2 View

Asking for a vector, containing Hygromycin B resistance gene?

In my work, Hygromycin B resistance gene is require for screening. I tried to amplify this gene from the vector pIJ941 but failed. I will be thankful, if anyone help me out from this problem by...

19 June 2024 7,821 3 View