Construction of pcdna3.i
WebMar 16, 2011 · The pcDNA3.1 vector encoding the wild-type Arf6 protein was obtained from the Missouri S&T cDNA Resource Center. pcDNA3.1-Arf6 T27N, pcDNA3.1-Arf6 T44N, pcDNA3.1-Arf6 Q67L, pcDNA3.1-Arf6 T157A, pcDNA3.1-Arf6 G2A/Q67L, and pcDNA3.1-Arf6 G2A/T157A mutants were created by a two step PCR with the first step introducing … WebOct 26, 2015 · After digesting the PCR product and the plasmid, the cfp10 fragment was ligated into the vector pcDNA3.1 (+). Correct insertion was confirmed by colony PCR, restriction enzyme digestion, and sequencing. Results: Electrophoresis of the PCR product on gel showed a 303-bp target fragment.
Construction of pcdna3.i
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WebMar 8, 2024 · Plasmid construction. pcDNA3.1 (+) vector was purchased from Thermo Fisher Scientific. STXBP5-AS1 DNA was obtained by PCR using cDNA, Pfu DNA polymerase and synthetic oligonucleotide primers incorporating restriction sites. PCR products were ligated into the pcDNA3.1 (+) vector according to the manufacturer’s … WebPlasmid Construction. pCDNA3-FLAG-PLAG1 (=pKH26) was constructed by ligating in frame the MscI/XhoI fragment isolated from the pCDNA3-PLAG1 expression construct (4) into pCDNA3.1FLAG (a kind gift of Stefan Pype of the laboratory of Cell Growth, Differentiation and Development, KUL, VIB). This enables
WebMar 14, 2024 · We provide a detailed, step-by-step protocol for donor vector design and construction using the pcDNA3 vector . Custom donor vectors can be difficult to clone and expensive to purchase. We provide a simple, efficient protocol to obtain custom donor vectors from the common pcDNA3 mammalian expression vector. WebpcDNA3.1 (-) CMV. CD was successfully constructed and CD-expressing Hep-2 cells can be killed by 5-FC, so the recombinant plasmid may be a candidate vector for laryngeal cancer therapy. [A study on construction of plasmid pcDNA3.1 (-) CMV.CD and transfection into laryngeal cancer cell Hep-2] Lin Chuang Er Bi Yan Hou Ke Za Zhi.
WebMar 14, 2024 · We provide a detailed protocol on how to design and construct a custom donor vector using Gibson assembly (and the commonly utilized pcDNA3 vector as the backbone) allowing researchers to obtain specific gene modifications of interest (gene truncation, gene deletion, epitope tagging or knock-in mutation). WebMethods: A bicistronic vector pcDNA3.0BA was constructed from pcDNA3.0. HCV PC154 gene and HBV preS2S gene were inserted into this vector to form bicistronic expression construct pcDNA3.0BAPC154S2S and monocistronic expression construct pcDNA3.0BAPC154 or pcDNA3.0BAS2S.
WebJun 15, 2015 · Construction of recombinant pcDNA3-HBsAg-p30-ROP Fresh recombinant plasmid pcDNA3-p30-ROP2 was extracted. Restriction enzyme digestion was performed …
WebMar 14, 2024 · We provide a detailed protocol on how to design and construct a custom donor vector using Gibson assembly (and the commonly utilized pcDNA3 vector as the backbone) allowing researchers to obtain specific gene modifications of interest (gene truncation, gene deletion, epitope tagging or knock-in mutation). lord and taylor bill paymentWebconstruction pcDNA3.1 vector and the PCR product of IL-33 were double digested by EcoRI and NotI and Figure 2. IL-33 and Fra-1 protein expression after pcDNA-IL33 transfection. *P < 0.05, compared with control. Figure 3. Transwell boyden chamber detection of IL-33 impact on cell inva-sion. *P < 0.05, compared with control. BioRad … lord and taylor bill pay loginWebOct 19, 2024 · The following vectors were used for construction: pcDNA3.1(+) (Invitrogen, United States), pGL4.10[luc2] (Promega), and pEGFP-C3 (Clontech, United States). The pRL-tk (Promega) plasmid was used to normalize luciferase activity. Plasmid DNA was purified with a Plasmid Miniprep kit. The DNA fragments were isolated from agarose gel … lord and taylor birdcage restaurantWebWe present a detailed method for constructing a mammalian cell-based full-length antibody display library for targeting hepatocellular carcinoma. Two novel mammalian library … horizon bcbsnj telehealth policyWebHence, a DNA vaccine, named pCDNA3.1 (+)-MCP-Flag, was constructed by inserting the cloned LMBV major capsid protein (MCP) gene into the pCDNA3.1 (+)-Flag plasmid. The expression of the recombinant plasmid was confirmed by Western blot (WB) and RT-PCR. horizon bcbsnj state employeesWebOct 26, 2015 · Mycobacterium tuberculosis is an etiological agent of human tuberculosis (TB). Designing new vaccines, including DNA vaccines, may be a useful strategy for … horizon bcbsnj refund addressWebContents: First plasmid construction：pCDNA3.0-PGK-BSD Annotation：pCDNA3.0 is a backbone plasmid ideal for transient expression in our designated cell lines including HEK293T,HepG2 and Bocs. All elements of our project are first to be incised into pCDNA3.0 and tested during transient expression in cell lines. lord and taylor black dresses racks