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
Old in-situ hybridization protocols require murine tissues to be fixed with PFA pH9.5, but I was wondering if the newer RNAscope technnology allowed us to work with tissues fixed at physiological...
29 March 2023 2,284 1 View
In my experiment to study the migration of microglial cells, I employ scratch assay/wound healing assay. I wonder if it is compulsory to use Mitomycin C to ensure the scratch covered by the cell...
29 March 2023 8,035 3 View
Hi everyone! I am expressing two 250kDA fusion proteins in E.coli BL21. They have been codon optimized (once) for E.coli expression but it seems that we can't purify only one band. Even after...
28 March 2023 8,475 5 View
Hello! I have a question about the pores in membranes. Membranes made of nitrocellulose, PVDF, nylon and other polymers are widely used in laboratories for blotting, filtering, etc. Almost any...
28 March 2023 9,783 3 View
Hello everyone, I have one doubt regarding cloning ? I am using pET30a (+) vector for cloning (5422bp) and Insert size 804bp. I tried all the ligation reaction ratio lilke 1:3, 1:5, 1;7 (100ng...
27 March 2023 6,985 6 View
Hi, I'm using A549 and PR8 H1N1 virus to do plaque assay and I'm facing a problem as my cells always wash off after I fix it using 4% formaldehyde. Currently, I'm using 2ug/ml TPCK, 0.6% agarose...
24 March 2023 813 0 View
Hello, I am having trouble with transient transduction of lentivirus particles in THP-1 cells and differentiated macrophages. I do not have an antibiotic resistance gene in my vector to do a...
24 March 2023 8,709 2 View
Can I purify the human rad54 protein using Bacterial Expression System? So far I just see any publication about rad54 protein using Insect system.
24 March 2023 9,784 0 View
I am trying to use overlap extension PCR to combine 2 linear PCR fragments around 1kb each. I amplified both fragments with overhanging primers with a 20 bp overlap between the two fragments. When...
23 March 2023 9,895 3 View
i need to find the common regions between them to make a tree that describes the relation between them or may i use specific gp of genes in the comparison instead of the whole genome?
23 March 2023 2,858 2 View
Hi all, I have to postpone some stainings and imaging due to technical difficulties, but the cells are already in culture. They are hiPSC derived cortical neurons in 96well plates and the staining...
23 March 2023 4,577 3 View
I have isolated the Triton insoluble protein aggregates from the cell culture media and blood serum. I got the pellet (insoluble protein) in the final stage of the isolation process and it’s not...
22 March 2023 9,941 10 View
I’m doing a cartwheel assay to see if a bacterial strain I’ve deemed resistant from an MIC has efflux pumps. Would it be possible to use Nile red instead of EtBr for the cartwheel assays instead?...
21 March 2023 7,112 1 View
Hello! I’m going to work with toxin/antitoxin selection system (eg StabyClone). I want to prepare electrocompetent cells with ccdB gene in genome.
21 March 2023 2,584 3 View
How do I know if my in situ hybridization probes binds mRNA and not genomic locus?
19 March 2023 2,035 7 View
Anyone with experience with cloning using E.coli DH5Alfa and Pjet 1.2 plasmid who can share their experience with me? I've been trying to understand and identify if my clones are showing DNA...
17 March 2023 8,509 5 View
I am getting 3PCR products of different sizes when amplifying off a plasmid, and I think it is due to unintentional primer binding. I am amplifying a trimer off the plasmid, and the 3 sizes...
17 March 2023 8,134 1 View
Hi all, our lab is currently working on H&E staining with frozen liver section. As you may see, the image shows that our tissue section has multiple large cracks. When the liver was...
16 March 2023 4,628 3 View
Hello How to extract DNA from water samples without using filter to perform metagenomics?
16 March 2023 2,299 3 View
What is the process/ protocols/ fundings available for Post doctoral fellow position in Pharmaceutical chemistry?
15 March 2023 7,243 1 View
I performed exosome isolation from cell culture media by using a kit for cell culture media. Now, I would like to perform the isolation of exosomes inside the cells. Which protocol can I follow?
15 March 2023 4,588 4 View
Hi, I am trying to extract cytosol-enriched and nuclear-enriched fractions from cultured cells. For the cytoplasmic extraction, I am using a hypotonic lysis buffer that contains DTT...
14 March 2023 9,323 2 View
Hello, I use to perform maxiprep but recently I have had a problem while I check the DNA through a digestion in agarose gel. I transform the plasmid in bacteria JM109, then I put a clone in a...
14 March 2023 6,644 2 View
Hello everyone, Does anybody have experience with electroporation of Cholangiocarcinoma cell lines? I was trying to find published settings for a pilot experiment involving the electroporation of...
14 March 2023 1,948 0 View