JRD247 Stable Transfection Cell Strain Construction Experiment
stable expression of cell line (steady transfer Cell strains refers to the continuous stability expression of a particular gene or interfere with specific gene expression in cells. The purpose of steady cell strain is integrated into the cell chromosome, allowing the cells to express the gene long term.
In the study of gene function, it is an effective transfection of target cells and is one of the prerequisites of functional research. Steady expression of cell lines make up for the transient infection (or transfection) in some functional experiments due to short defects in exogenous genes, facilitating long-term observation genes for cellular function and inter-protein interaction. The method of introducing a target gene-containing carrier is introduced into the cell using a method of transfection plasmid or virus into the cell, and the respective drugs are selected for screening mixing positive cloning according to the resistance marks contained in different gene carriers. Based on positive mixing clones, positive monoclons having a single cell length were obtained, and the cell line was stably transfected. Common eukaryotic expression vector resistant markers have puromycin, hygromycin and neomromycin.
Shanghai creature has been experienced with steady cell strains for many years, which has been in H1299, THP-1, MCF7 / ADR, A549, V79, MC3T3-E1, CHO, SiHA, 3D4 / 2, HL7702, SW480, Hepg2, SGC-7901, MGC-803, HT-29, SAOS-2, U-2OS, PC12, SK-BR-3, MCF-7, HMSC-AD Gist-T1, HEK293, 293T, etc. Overgraduated and RNAi steady transcetum strains were successfully constructed in cells.
Product Features:
1, Stable: 15 generations Stabilized transfection cell stable expression.
2, Quick: General control is completed within 1 month.
3, economy: affordable, cost-effective.
4, quality: Strict identification of exogenous genes.
Two methods of stabilizing transfection cell line construction experiment
1, after transfection plasmid, the stabilized cell line was screened. Lower transfection in most cells. For gene over expression, if the transfection efficiency reaches 40%, the gene overgraduates can be expressed. However, for gene interference experiments, the transfection efficiency is much higher than that of gene overcurrent, and usually plasmid transfections cannot be satisfied. In addition, the plasmid travels in the cytoplasm, quickly degraded, can not meet longer detection items; the level of plasmid transfection is extremely low, the stable cell line constructs consumption, and the rate is very low, often need to pick monoclonal Strain. 2, viral infection screening stable cell line viral infection method Compared to plasmid transfection screening monoclonal method is more convenient and efficient, is current mainstream Stable cell line screening method. It is an ideal vector for preparation of stable cell lines due to its high-efficiency integration, high efficiency transcription, high efficiency expression, high-efficiency, high infection efficiency, and integration with cell chromosomes without gene rearrangement.
Stable expression of cell line construction service process 1, vector construction: Construction of exogenous genes to a suitable carrier, if virus transfection, packaging Virus.
3, Cell inoculation: Transfection Experiment The daytime in breeding cells, the flat density of various cells is determined according to the growth rate of various cells and the shape of the cell. The transfection was carried out to achieve 60% -80% 4, cell transfection: transfected with the constructed carrier (or packaging virus). 5, antibiotic screening. 6, identification screening results. customer needs
1, construct a good foreign gene carrier or genetical template.
2, cell line to be transfected( 1 u0026 Times; 106 cells, capable, multiplier, less than 48 hour), special cells need to provide medium. We offer 1, construct a good foreign gene carrier. 2, construct a good stable cell line. 3, constructing a stable transform cell line experiment report and related exogenous gene expression analysis. Currently, AY Pharma, Main Experimental Technical Services, Service, Animal Model Construction, Cell Strain, ELISA Kit, Welcome to the majority of scientific research enthusiasts to come to consult!