I used the cas9 to KO my gene in HEK 293T cell , but one of my control negative cell lines show the increasing of my target protein. I didn't transfect an extra homologous sequence. Seeking for guidance .thanks
03 March 2021 8,272 1 View
Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...
03 March 2021 4,275 4 View
Anyone can recommend any books on such topics? Thanks in advance.
02 March 2021 8,457 2 View
I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...
28 February 2021 5,440 3 View
Recently I face some problems with the mathematical ( calculation) on the reference governor and command governor, so I need some help from you all to get which platform or journal can let me...
23 February 2021 7,746 2 View
According to the Sentinel-2 user guide here (https://sentinel.esa.int/web/sentinel/user-guides/sentinel-2-msi/naming-convention), the compact naming convention is arranged as...
21 February 2021 5,919 9 View
For example, I have this phonon dispersion(Fig. 1), and now I would like get vector of atoms which describes TA1 in P. I know v_sim can show vibrational animation of specific mode, but I do not...
20 February 2021 4,332 4 View
20 February 2021 5,984 3 View
I was analyzing some metabolites in human plasma. While the intraday precision of these metabolites was acceptable ( less than 20%), the interday precision was quite large. So what influences...
13 February 2021 9,183 9 View
When co-transfect S2 cell with selection vector (pCoHygro or pCoBlast ) to make a stable cell line, does the gene integrate into the genome or just like E.coli? How do the plasmids amplify within...
09 February 2021 5,423 1 View
Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...
01 March 2021 210 1 View
There are many ways for mRNA to intervene with genome expression. This question is raised for experts in CRISPR systems. Any RNA does not stand alone within acell. There are many ways to become...
18 February 2021 9,109 5 View
I want to completely delete or largely truncate a gene and not only achieve a functional knockout in a cell line. I can only transfect my cells with the Crispr/Cas9 sgRNA RNP and not introduce a...
08 February 2021 2,032 2 View
I am working with myelin-related proteins and interested to do cell experiments with my target protein in cells. I have created lentiviruses for my proteins, and mutants and also CRISPR set-up for...
02 February 2021 8,083 1 View
Hello. I have a viral plasmid which contains a bicistronic region: CMVprmtr-GFP-IRES-PuroR. I would like to cut out the GFP sequence to get rid of the green fluorescence. In this case, would it be...
02 February 2021 1,644 7 View
Hello, I hope to insert a gene into mammalian cells in culture using CRISPR/Cas9 and an HDR template. Will insertion of a 1 kilobase gene cause chromosomal abnormalities and effect cell...
31 January 2021 2,516 1 View
Hi Everyone: I have a question regarding to qPCR. I am trying to do RT-PCR, and I want to have equal amount of input DNA for my RT-PCR, so I normalize my DNA input based on qPCR values of a...
31 January 2021 6,977 4 View
Hi, I'm trying to create a stable overexpression line with EF1a-FlagGOI-IRES-GFP using lentivirus packaged with pxPAX2 and pM2G. After transduction of 293T with 8ug/ml polybrene, I was able to...
29 January 2021 1,878 4 View
I am optimizing a CRISPR/CAS9 system in vitro before using it in vivo. I would like to transfect AML12 cells with my pAAV-sgRNA-vector to validate editing efficiency before generating virus.
26 January 2021 8,733 2 View
Hello, I want to insert a gene into my cells using sgRNA, Cas9, and an HDR template. The gene that I will insert into has its own promoter. Because I will cut the gene downstream of its start...
14 January 2021 1,170 2 View