Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a... | Contact experts in Plasmids to get answers
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Questions related to Plasmids
I could not find a study that used CRISPR-mediated HDR for stable integration of large genes. I would be grateful if anyone could point me in the right direction.
06 August 2023 440 2 View
I am facing a problem while running BRIG with genbank files. I have given plasmid sequences as my input files. I want to give gene annotations & following the manual I have added the genes...
06 August 2023 693 0 View
Does it have something to do with the concentration?
04 August 2023 1,515 3 View
I am starting my CHIP qPCR on MCF7 and MDAmb231 cell lines and I am Using the IMPRINT CHIP kit by Sigma. I need to optimize the sonication time for both cell lines, but the buffers are very...
04 August 2023 3,161 0 View
I used 0.5 ug plasmid. Cell confluency around 60%. Transfection time is 48 hour. I used empty plasmid (pCMV-3Tag-4A) and only Lipofectamine as a negative control. However, I used EGFP plasmid tag...
04 August 2023 1,831 2 View
Why do we add media first and DMSO later?
03 August 2023 3,136 6 View
I want to transfect mRNA in 3D cardiac models. Assays using Lipofectamine or iTOP didn't work, and I didn't find any method that efficiently reaches the center of 3D models. Thank you for your help!
01 August 2023 5,271 0 View
I need to prepare conditioned medium for Jurkat cells. I understand that it is medium in which the cells have already grown, and the cells filtered out, but how exactly should it be made? What...
01 August 2023 6,371 4 View
I am seeking to understand the variations in molecular mechanisms between pathogens involved in sepsis/sepsis-induced DIC and SARS-CoV-2. Additionally, I am interested in exploring how these...
31 July 2023 4,400 2 View
Hello. We did golden gate assembly where in part vectors we chose kanamycin resistance gene and in destination vector we chose ampicillin resistance. Why is that?
31 July 2023 5,444 3 View
I have been growing multiple Vibrio species in the lab continuously with no problem. Today after checking plates that are 2 weeks old, I noticed these strange track marks all over the plate. It...
31 July 2023 6,014 2 View
I am facing a problem to circularize a particular bacterial plasmid sequence while doing hybrid assembly in unicycler. After the assembly, it's not showing circular. While the other plasmids have...
30 July 2023 1,239 1 View
The CSI phylogeny 1.4 no longer allows to build a phylogeny with more than 50 isolates.
29 July 2023 7,609 5 View
Anyone know the problem? why the Ct values for my samples are fluctuating so much and even not reaching threshold line? is this related to my unspecified primer? because after I checked my primer...
28 July 2023 8,553 9 View
I’m trying to reproduce the library transformation efficiencies seen in this 2010 paper: An improved yeast transformation method for the generation o... in which they claim 1.5 x 10^8 cf/ug DNA....
27 July 2023 4,134 3 View
I want to use yeast as an expression system. So, I wanted to ask if Debaryomyces hansenii could be used for the same. Although, Saccharomyces is a model system for gene expression studies. Can we...
27 July 2023 3,719 2 View
Despite finding many websites for primer design in promoter regions, I am in need of guidance from someone experienced to direct me through the appropriate steps.
25 July 2023 9,914 5 View
Hello, I am currently working on generating deletional mutants in Agrobacterium C58 using a construct containing upstream and downstream fragments of a target gene. The construct also includes a...
25 July 2023 8,442 2 View
Hi everyone I inserted a restriction site (5 nucleotides) between RBS and the start codon (these 5 nucleotides are upstream of multiple cloning site). after confirming my cloning, we started...
23 July 2023 1,339 5 View
Want to know how transfection of mutants in cell lines can be performed in effective manner? Any detailed protocol which would be cheap and easy to perform kindly suggest.
23 July 2023 8,005 3 View
Chikungunya virus, C6/36 cell, morphological changes
22 July 2023 9,802 2 View
Could the lower transformation efficency be problematic in practice?
21 July 2023 3,469 3 View
I am trying to express a novel GlcNAcT enzyme in BL21(DE3). I have successfully cloned it in pET28a vector and confirmed it through sequencing. I tried following previous protocols for GlcNAcT...
20 July 2023 6,052 2 View
Please advise on this. I have tried all sorts of troubleshooting like- pH adjustment, ventilation, inversion of the tube gently after adding P2 buffer, overnight culturing time, optimizing the...
20 July 2023 5,485 2 View