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
Elution buffer composition is- 50mM NaH2Po4, 300mM NaCl, 250mM immidazole at pH 8. PI of my protein is 6.7. I purified My protein using Ni-NTA resin and run sds page and till the completion of gel...
24 February 2023 4,645 12 View
I am working on the transfection of a murine cell line. I have to transfect 4 individual genes, so I have 4 individual plasmids, each with a different antibiotic resistance gene. Before...
24 February 2023 7,191 2 View
Actually, I'm working on an RNA extraction kit, but due to an unknown reason, I'm not getting the desired result. Although, some bands appear on MOPS-Formaldehyde gel (smear-like) but didn't get...
23 February 2023 954 3 View
We work on primary Cancer cell derivation from patient samples so i want to immortalize a cell type can anyone suggest lentriviral protocol
23 February 2023 6,702 2 View
Hi all, Can someone help me understand the difference (if there is one!) between an enzyme's substrate specificity (kcat/KM) and its specific activity towards different substrates? If I want to...
23 February 2023 8,333 1 View
Hi everyone, I performing a site directed mutagenesis on a wildtype plasmid and I have been having troubles generating the mutant plasmid I want. I am hopping to make a post here and see if...
22 February 2023 1,320 5 View
I am using thermofisher midiprep kit for my plasmid isolation. I got 43ng/uL last year. However, I'm using the same protocol now and getting consistently lower yield (around 7ng/uL) in 350 uL EB....
22 February 2023 9,843 3 View
Some of my backbone expression vector are clearly contaminated by this IS. I use DH5alpha or TOP10 for routine molecular biology and quite surprise to observe this type of insertions in...
21 February 2023 3,294 3 View
I'd like to know What is the minimum and maximum amount of Plasmid DNA that can be used for transformation of Bacteria ?
20 February 2023 9,797 3 View
Is this safe for the RNA probe?
20 February 2023 2,585 2 View
Is there any explanation for strong, adequate or low value of it? Thank you
20 February 2023 2,517 4 View
Bacteria culturing is an essential part in the medical field especially in identifying treatments such as antibiotic prescription for a certain disease. In addition to this, bacteria culturing and...
19 February 2023 5,751 3 View
Dear colleagues, I am trying to figure out the principle of manually designing gRNA in order to clone it into plasmid. I know, that I have to add sticky ends to my oligos to make cloning them...
19 February 2023 755 2 View
What is the protocol for lipid stain in HepG2 cell line using Oil Red O stain?
19 February 2023 7,646 1 View
I am cloning five genes of approx. 1.2 kb each into pET-28a(+) vector (separately, i.e., not all in the same vector) by the USER method. The protocol is essentially the same for all constructs,...
18 February 2023 5,058 8 View
Dear all, I have transformed ZYCY10P3S2T E. coli (used for minicircle production, should be recA-) with following ligation mixture: 1. Insert: 3353 bp sequence containing gene of interest 2....
18 February 2023 9,634 4 View
my protein is 151 kDa, but its Instability index over 80, so it may be unstable in buffer.
17 February 2023 9,666 0 View
I will be performing both Sanger sequencing and RT-qpcr for my target gene. For Sanger, I have ordered 3 sets of primers (around 2500, full length). Is it possible to use the same primers for...
16 February 2023 3,624 3 View
Greetings, I am currently working on a project that involves examining the differences in gene expression within groups. During the course of my research, I have discovered that the primers I am...
15 February 2023 2,908 2 View
reverse genetics platform is used for cloning platform where cDNA is made from Viral RNA and is used for cloning
15 February 2023 4,615 0 View
Hello, I am trying to do IVT from cDNA. However, I do not get any RNA! I have included the control RNA sample that comes with the kit (Message Amp II) and that does not work either. 260/230 ration...
15 February 2023 4,721 8 View
Hello , I am considering using the XTEN linker (SESATPES) in a Cas9 fusion protein I may later want to sell. I´ve seen XTEN refered to as Amunix XTEN. I´ve read some patents about XTEN like these...
14 February 2023 2,739 1 View
I am trying to clone with a pMAL-C2X vector carrying BamHI and SalI sites. I performed several times the ligation with the PCR insert and found several colonies. But seems all are empty vectors....
14 February 2023 3,947 3 View
I am trying to replace a 20 bp region with a different sequence of same lengthin a 10kb plasmid. I designed overlapping primers. However after PCR I am only getting the template. At first I...
14 February 2023 3,636 3 View