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Questions related to Cloning
Dear all, I'm planning for continuous overexpression and Knockdown. I'm using Cre-ERT2 system, and i want to firstly overexpress YAP and then knockout YAP in lung. i have made loxp-YAP-loxp, so...
21 September 2020 9,629 3 View
Hi! I'm wondering if anyone else has had a situation where they are purifying an enzyme, and only one clone is active? The cloned gene is genetically identical to other clones, but only this one...
19 September 2020 8,098 3 View
I am trying to generate monoclonal antibody by phage display method. After mRNA to cDNA conversion, how many numbers of cDNA from the cDNA pool to be cloned into phagemid vector. What determines...
18 September 2020 9,428 4 View
I am cloning a whole cDNA of a particular gene and I have designed forward primer at start of 1st exon. The region downstream of forward primer is extremely high GC rich (around 85-89%). Because...
18 September 2020 6,647 11 View
Hello all, I will do oligonucleotides annealing for the first time which will be later used for cloning without any more modification. Can any of you kindly suggest if I should purify the annealed...
17 September 2020 4,007 3 View
Hi all, I am new here and I am about six weeks in to my first ever research project! (I just came here to kick-start my career as a researcher.) :) The aim of my research project: "The aim...
16 September 2020 9,968 3 View
Some triploid acacia clones in Viet Nam have shown high potential in growth, disease resistance and good adaptation.
15 September 2020 2,038 3 View
Hello I work with Roche RACE PCR kit and I have a problem with 5` RACE PCR. When I checked my PCR product on Agarose gel, I only saw an unspecific band. I`m not sure about dA-tailing step,...
14 September 2020 9,417 0 View
After I have cloned my gRNAs into the lentiCRISPRv2 vector and sent the construct for sequencing, I have noticed that a single mismatch between my clone and original sequence has been introduced....
12 September 2020 4,185 1 View
Hi, I generally made LV in HEK293T cell lines by triple transfection. I am wondering whether the newly generated LV will transduce the packaging HEK293T. In this way I could exploit the...
10 September 2020 6,445 3 View
My idea is comparing the S gene (>4kb) of the new strain of PED virus with the novel strain (sequence already available on GeneBank). Most of the papers I have reviewed were cloning the S gene to...
08 September 2020 944 12 View
I am trying to clone using the pcDNA3.1 V5-His-TOPO expression kit. There is no problem transferring it to the cell, but I can only transfer the vector. I did not succeed in vector-insert...
07 September 2020 3,571 0 View
Dear members! I have two questions/issues: 1) Does anybody know how to solve the problem with FD Enzymes for preparation of cloning vectors? I make restriction before cloning with FD XbaI + FD...
03 September 2020 2,602 4 View
maybe someone with experience in the field of cytotoxicity testing of materials for medical use may help me. I have two possible cell lines available in my lab: mrc-5 and 3T3 fibroblasts....
02 September 2020 6,097 4 View
I need to do isotyping on some purified rat monoclonal antibodies. The isotyping is done using in-house light/heavy chain coated on ELISA plates. Plates were blocked with 10% CBS overnight. The...
01 September 2020 1,010 2 View
I am checking the additive, synergistic and antagonistic effects of mixed chemicals based on the EC value and other physiological parameters. I read some papers using different softwares such as...
21 August 2020 5,938 5 View
A normal vector can withhold a gene of insert upto a capacity of 10KB.What if we were to insert a gene of 60KB?
17 August 2020 6,920 1 View
Dear colleagues, I am looking for a python library equivalent to the one found in the bioconductor annotation package for TxDb object "TxDb.Hsapiens.UCSC.hg19.knownGene". Also, I am interested to...
16 August 2020 4,222 3 View
1- In a typical mammalian cell, the protein content is 20 % of its net weight. If the density and volume of the cell are 1.2 g/mL and 4 × 10-9 mL, respectively, then the concentration (in mg/mL)...
12 August 2020 3,643 3 View
Hello, I am new to molecular cloning and I have been working on cloning my passage samples for almost a month without success. I was able to successfully clone my wildtype but it is incredibly...
04 August 2020 5,781 3 View
I digested my lenti guide puro plasmid. The only modification in this plasmid is that it has blasticidin resistance gene instead of puromycin. I get the desired bands at ~8kb but I do not see the...
04 August 2020 2,932 4 View
I am trying to blunt clone a 1 kb PCR fragment into pBluescript. The PCR product has restriction sites at the ends, but I have never had much luck trying to cut ends of PCR products and clone...
02 August 2020 2,692 8 View
Dear researchers, I have one query about the evaluation o drug release for the ointment. We want to investigate the drug release profile of ointment. but we do not have scientific instruments like...
30 July 2020 8,394 4 View
This can refer to 2 different journals with the same details
25 July 2020 9,880 7 View