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Questions related to Cloning
Dear, I'm developing an in-house indirect ELISA to detect the present of antibodies in the serum against target protein of the pathogen. The setting of my ELISA is that I have the target protein...
04 September 2023 4,303 7 View
Gene around 5000bp
01 September 2023 9,935 3 View
Hello, I am looking for lac deficient strain E. coli BL21(DE3) for preliminary screening of clones carrying a beta-galactosidase gene. Would someone be willing to share the strain or direct me to...
01 September 2023 870 0 View
Dear colleagues I had a Prism file containing data until a few days ago; it opened without any problems Now it gives an error and does not open. Can anyone help me? Thank you
01 September 2023 8,815 1 View
Water-in-oil droplets are generated using Bio-rad droplet generator, and the oil phase is fluorosurfactant synthesised by myself dissolved in HFE-7500 with 2% W/W concentration. The water phase is...
30 August 2023 6,775 0 View
To check the expression of JAK2, STAT3, SRC and indoleamine 2 and 3 dioxygenase genes in the blood samples of people who do not have any immunological or hematological diseases, is the best way to...
29 August 2023 5,471 2 View
I have an insert size of 900 bp, trying to clone it in pET28a vector. But after every the transformation when i am doing colony pcr and digestion check i can only get the fragment of 500 bp. Why...
28 August 2023 2,416 3 View
Nuclear tracks in solids or SSNTDSs Or NTDs
27 August 2023 9,920 3 View
I am doing overlap PCR to join 5 fragents to form expression cassette and product is always trapped in the wells and is not migrating down, I did purification by PCR buffer A but still the result...
23 August 2023 5,345 9 View
Hello, I've been having issues with my cDNA PCR which used to work previously. I'm working with cDNA from velvet worms, amplifying Cox1. Earlier, I had successfully amplified Cox1 with specific...
23 August 2023 6,259 4 View
Dear Researcher, Different software tools can produce maps similar to the ones above. How can I find the best software for making genome maps like this?
23 August 2023 1,523 1 View
Which tissue is responsible for flexibility in plants and which cambium is responsible for the secondary growth is present in this stem?
22 August 2023 4,790 3 View
Hello I am doing an eQTL study and here are the details for that. I have two mouse strains at two different time points. I already have extracted the RNA fro my tissue of interest and got done...
21 August 2023 1,558 1 View
We have been managing a complex case, an 8yo Staffordshire Bull terrier with several cancers including a lung tumour thought to be a possible biphasic pulmonary blastoma; the epithelial cell...
21 August 2023 5,471 0 View
I am growing some leukemia cell lines (MV411, CEM) and suspect that I might have mycoplasma contamination. I see black dots in my cell culture, and I am unsure if it is cell debris or mycoplasma....
17 August 2023 3,207 5 View
HepG2 was fine until last Saturday(third picture), but today they detached and the color turned black...(first and second picture) Are they dead? or contaminated? After the subculture, I could...
16 August 2023 6,037 5 View
Hi, I'm prepping a sample for Oxford Nanopore GRIDion sequencing using the old PCR-cDNA Barcoding kit (SQK-PCB109). The protocol specifies Agencourt AMPure XP beads should be used for purifying...
14 August 2023 989 0 View
I am trying to optimize a SYBR assay and one primer set is giving me dimers every time. I've increased the annealing temperature to close to the calculated value for that primer set, and the other...
10 August 2023 2,879 4 View
I was working on a gene construct synthesized in pet29 vector as a clone. Primers were prepared and optimized with gene at Tm 58 degrees. Once primers were optimized, I carried out transformation...
09 August 2023 5,647 1 View
in network mobile
09 August 2023 5,800 0 View
I am using a three vectors system to package lentivirus in 293T cell lines. I have inserted GFP into my transfection vector. And I transfected all three vectors into 293T cell line with PEI and...
08 August 2023 3,732 5 View
Is there any online tool available to check, if designed primers are suitable to get overexpression of the histidine-tagged protein, cloned in pET28a vector? How to confirm the suitability of...
08 August 2023 8,248 1 View
I'm trying to knock out a gene in E. coli BW25113 ΔclpS using the phage λ Red recombinase gene knockout method. I got a few kanamycin resistance colonies (confirmed by colony PCR) but without gene...
08 August 2023 7,575 0 View
I cloned a gene and the gene was confirmed by genomic complementation experiments. However, it can not be complemented while adding any tags. The molecular weight of the protein is very small,...
02 August 2023 8,481 0 View