Will the plasmid without SV40 Ori have a quite low expression level in 293T?

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How can we improve storage conditions of museum objects?

90% of museum objects worldwide are in storage.

21 July 2024 8,038 6 View

How to removal of dead cells and debris from organoid cultures?

I'm currently working with human pancreatic organoids and I am wondering to remove the dead cells and debris from health organoids, any suggestions would be appreciated!

17 July 2024 8,330 1 View

Does it work as antibiotic resistance if plamid has no RBS??

I am on gene work to express gene of interest in some Bacillus species. Thus far, i need to try some RBS in Vector which is optimized for bacillus subtilis. Before i design vector to express in...

11 July 2024 7,847 1 View

Can a liquid metal-ligand sample be directly analyze in flame atomic absorption spectrosocopy?

Can I directly analyze a liquid metal-ligand sample without acid digestion in flame atomic absorption spectroscopy?

25 June 2024 1,866 2 View

Is this a mixed methods approach?

For my PhD dissertation: If Chapter 1 is a systematic literature review (QUANTS/QUALS) while Chapters 2, 3, and 4 are empirical chapters (QUALS), is this considered a mixed methods research...

16 June 2024 3,220 5 View

What can I do if primer solution gives band in electrophoresis before PCR reaction?

Trouble 1: I ran a PCR reaction and found a low molecular weight band (100 bp) when doing electrophoresis. 1ng plasmid (OD260/280=1.84) was used as template. Final concentration of primers was...

13 June 2024 5,540 6 View

How to find the original questionnaire of the article-Dimensional analysis of schoolchildren’s food label comprehension: a pilot study?

27 May 2024 2,551 0 View

Why the Mixed mode simulation always exit with status 0?

I encountered some issues while simulating the transient characteristics of a device using Mixed Mode. After executing the following codes, an error message "Simulator has unexpectedly exited with...

23 May 2024 2,238 0 View

Can bands of cell lysate be seen during SDS-PAGE?

I want to detect my target protein through WB and it worked well before. But now I could not get any signal for both my target and the GAPDH, so I ran SDS-PAGE. I expected many bands on the gel of...

23 May 2024 613 6 View

Can I ask for a JCPDS card no. 01-083-1436 or CPDS file number 00-031-1979 for urea?

Can I ask for a JCPDS card no. 01-083-1436 or CPDS file number 00-031-1979? I need to index a urea derivative compound to the face-centered tetragonal structure, JCPDS card no....

15 May 2024 9,751 2 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

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 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

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 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 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

What is the best way to drop a plasmid?

Please address the best way to drop a plasmid. Background: I have a "bait" plasmid resistant to kanamycin and a "prey" plasmid resistant to carbenicillin. After many rounds of streaking on...

25 July 2024 2,532 3 View

Isolation of PLV6-Bmal-Luc plasmid?

I am currently at a stage in my research where I will be using the pLV6-Bmal-luc plasmid (Addgene #: 68833). However, I am encountering some issues with the isolation of this plasmid and am unable...

23 July 2024 5,296 0 View