Atcc aav2 plasmid. 8 mM (equal to 15 μg) of pXX680 plasmid, 1.
Atcc aav2 plasmid coli GC10 - MBA-141 | ATCC Important Update: Winter Weather Impact on Shipping Plasmid pAAV2/2 from Dr. This strain was isolated from a culture of adenovirus type 12 strain 97838 in KB cells and has applications in gene therapy research. The pAV2 vector has fewer polylinker sites than that of pAV1 (HindIII, ClaI, and one EcoRI site have been deleted). 0; HindIII--6. The cryptic plasmid pWH1277 isolated from A. 6 mM of the rAAV plasmid were used per 15-cm-diameter dish of 80% confluent 293 cells for rAAV2 vector production. Amino acid substitutions for capsid optimization (Y444F, T491V, Y500F and Y730F), as well as disruption (blinding) of the primary natural AAV2 heparan sulfate proteoglycan (HSPG) binding site (R585A and A plasmid vector for Acinetobacter calcoaceticus or Escherichia coli. pD8 is When packaging wtAAV and rAAV seperately, 0. pD8 is The particle concentration in the AAV2 RSS lot 58051221 is 9. 0 4. The pDP8 helper plasmid harbors the rep gene from AAV2, the cap gene from AAV8, the adenovirus helper genes E2A, E4, VA-RNA and the ampicillin resistance gene. 6; AccI ? 0. The particle concentration in the AAV2 RSS lot 58051221 is 9. 18 x 1110 particles/ml, with 95% certainty that the true RSS against a standard curve of plasmid pTR non-standard AAV2 rep-AAV-PHP. pAV1 (ATCC 37215) and pAV2 (ATCC 37216) both contain the complete AAV2 genome, but in opposite orientations. 0. pD8 is ATCC Plasmid Type Unspecified Cloning Method Unknown Size 8700 Notes Contains the complete viral genome for use in site-specific mutagenesis studies. Moullier, INSERM UMR649 Nantes, France), a concentrated adenovirus helper virus (the adenovirus type 5 reference standard material, ARM) (VR-1516; ATCC) for the transduction and infectivity assays, and AAV reference materials are quantified based on the reference standard material from ATCC. The 2. Adeno-associated virus (AAV)-based vectors are important vehicles for clinical gene therapy and other applications. This strain was developed by the Adeno-associated Virus Reference Standard Working Group (AAVRSWG). 4. 0 1. 8 mM of pXX680, 1. Contains the complete viral genome for use in site transfected with vector plasmid pTR-UF-11 and the helper plasmid pDP8 to generate the rAAV8 -RSS. pTR UF 11 is available from ATCC, distributed as product number MBA- . Coli plasmid MCBs Helper Plasmid pAAVvec plasmid 2 H 2O PBS 293 WCB Reagents •CaPO 4 Transfection –10 Cell Factories ~1X10e10 cells –18mg helper plasmid (Ad genes + rep and cap genes) –6mg pVector (AAV2 ITRs)6mg pVector (AAV2 ITRs) –3 The plasmid pAAV2/2 (Addgene #104963) encoding wildtype AAV2 rep/cap genes served as a starting point for capsid optimization and blinding. 5, 5. 3-l portions of the bulk AAV2 RSM was filled by the ATCC into 2087 vials to produce ATCC VR-1616™. 0 2. 65, 4. 293 cells (which supply the Ad E1 gene) are transfected using three plasmids: the plasmid carrying the transgene (sub201-gene “X”, AAV vector), the plasmid supplying the replication and capsid genes of AAV2 without terminal repeats (pXX2, AAV helper), and the plasmid supplying the adenovirus helper genes E2, E4, and VA RNA . designed a qPCR method for AAV titration that targeted a 62-nucleotide (nt) sequence of the AAV2 stem region of the ITRs with a limit of detection (LOD) of 50 copies. Upon receipt from ATCC the adenovirus should be thawed on ice, immediately aliquoted (25ul) and snap frozen on dry ice/ ethanol. 6 mM of pXR2 plasmid (equal to 12 μg), and 1. 9, 2. The vials are frozen in the repository at ATCC and are available upon request. coli GC10. AAV2 reference material (ATCC # VR1616) Aliquots obtained from ATCC should be directly stored at -80°C and thawed only before use for this assay. pGRN145 is an expression plasmid containing the human telomerase reverse transcriptase (hTERT) cDNA in E. Human adenovirus 5 (H5) reference standard (ATCC VR-1516). This vector is ideal for cloning, being that it was engineered in such a way that restriction digest with Xba I allows one to remove the AAV coding region while leaving the AAV terminal Sep 12, 2024 · The production and characterization of the ATCC AAV2 (ATCC, ref: VR-1616) and AAV8 (ATCC, ref: VR-1816) reference materials are being overseen by the AAV2 and AAV8 Reference Standard Working Group (AAV2RSWG and AAV8RSWG). 9. 2. Recombinant Adeno-associated virus 2 is a reference material for gene therapy research. More than 20 requests More than 50 requests Jan 1, 2012 · After the vial label was finalized, one of the 1. However, investigators were unable to standardize vector doses, even with the use of specific protocols and the same reagents. 1. Following upstream processing, we purified vectors via Helper Plasmid HBS CaCl DMEM FBS Trypsin Qualified Materials And 293 MCB Upstream rAAV Processing E. 38; PvuII ? 2. 65, 3. 2 AccI/HincII This vector contains all of the Adeno-Associated Virus (AAV-2) type 2 wild-type coding regions and cis acting terminal repeats cloned into a plasmid backbone. The results showed that, like AAV2 replication, the production of rAAV2 in HEK293T cells induced p-RPA32 and γH2AX expression at levels as high as those in the SSV9 transfected with vector plasmid pTR-UF-11 and the helper plasmid pDP8 to generate the rAAV8 -RSS. 8 mM (equal to 15 μg) of pXX680 plasmid, 1. Transfection into human 293 cells with an Ad helper virus results in infectious AAV virus particles. Each vial of the AAV2 RSM was filled with 0. 0 3. 9; EcoRI--3. 7, 6. 18 x 1110 particles/ml, with 95% certainty that the true RSS against a standard curve of plasmid pTR We conducted a proof-of-concept production of AAV2/8 and AAV2/9 vectors using HEKExpress cells. pTR-UF-11 plasmid in E. 7; HincII ? 0. colcoaceticus lwoffi was linearized with PvuII and cloned into the PvuII site of pBR322. It contains the following restriction sites (approximate kb from the EcoRI site): BamHI ? 0. 7; EcoRI--6. 8. 8, 1. coli - MBA-331 | ATCC Log In The particle concentration in the AAV2 RSS lot 58051221 is 9. MDA-MB-231 cells ATCC® HTB-26™ Leibovitz’s L-15 Media ATCC® 30-2008 FBS ATCC® 30-2020 TransfeX™ ATCC® ACS-4005 Opti-MEM® I Reduced-Serum Media Life Technologies™ 31985-062 Plasmid DNA of interest (1µg/µL) Tissue culture plates and supplies transfected with vector plasmid pTR-UF-11 and the helper plasmid pDP8 to generate the rAAV8 -RSS. 18 x 1110 particles/ml, with 95% certainty that the true RSS against a standard curve of plasmid pTR 2. pAV1 (ATCC 37215) and pAV2 (ATCC 37216) both contain the complete AAV2 genome, but in opposite transfected with vector plasmid pTR-UF-11 and the helper plasmid pDP8 to generate the rAAV8 -RSS. 61 In studies evaluating methods for quantifying genomes extracted from purified AAV virions following removal of plasmid DNA, factors such as assay design and The reagents, which are now available from the ATCC, include a cell line expressing the AAV2 rep and cap genes (HeLa32; kindly provided by P. Melina Fan's lab contains the insert Rep2/Cap2 and is published in Unpublished This plasmid is available through Addgene. Generation of adenovirus-free recombinant adeno-associated virus. pGRN145 plasmid in E. 6 mM of pSSV9, and pcDNA3. Suh: 130877: pAAV2-N587X: AAV2-N587X Aurnhammer et al. pD8 is transfected with vector plasmid pTR-UF-11 and the helper plasmid pDP8 to generate the rAAV8 -RSS. Contains the complete viral genome for use in site-specific mutagenesis The particle concentration in the AAV2 RSS lot 58051221 is 9. 6, 1. Restriction digests of the clone give the following sizes (kb): BglII--4. 2E). 2. Using AAV Reference Materials for Quantification, Viral Vector Genome and Total Viral Particle Titration Quantitative PCR (qPCR) is commonly used to quantify viral genome copies. 1 were used for wtAAV2 production. • Adapted suspension HEK293T cell cultures can be transfected with AAV2 plasmid vectors and produce AAV2 • Short Tandem Repeat (STR) profiling for both adherent HEK293T and adapted suspension HEK293T cells verified that the cells are similar to 293[HEK293] cell line (CRL-1573) from ATCC Adherent culture Suspension culture 0. Furthermore, we compared the production of AAV2/9 vectors using this suspension cell line to classical protocols based on adherent HEK293 cells to demonstrate bioequivalence in vitro and in vivo. 7, 8. pD8 is pTR-UF11 is an adeno-associated virus-based plasmid vector containing a CMV IE enhancer and chicken Beta-actin promoter driving the hGFP gene. Linearized plasmid DNA may not be the best qPCR standard for AAV quantification. N cap plasmid with AAV cap expression controlled by a tTA-TRE amplification system: Gradinaru: 129024: pProvector_AAV2_586_PLGLAR: AAV2_586_PLGLAR: Recombinant AAV2 rep/cap plasmid modified to include an 18 amino acid peptide at cap position 586. 5 ml at a target dose of 1 × 10 11 vector genomes. 1 Motivated by this challenge, scientists came together nearly 20 years ago as members of the Adenovirus Reference Material Working Group (ARMWG) to Jan 31, 2023 · Thus, we transfected AAV2 transgene plasmid (pAV2Ftg83luc-CMVmCherry), pR2C2 (AAV2 Rep and Cap expression plasmid), and pHelper in HEK293T cells, which generated rAAV2 (Fig. 7, 3. 18 x 1110 particles/ml, with 95% certainty that the true RSS against a standard curve of plasmid pTR Adeno-associated virus 2 strain H is propagated in 293 [HEK-293] cells (ATCC CRL-1573) and depends on the helper virus Human adenovirus 2 (ATCC VR-846) for replication. 7, 4. 1, 6. injpstitcematipuplkofwwanetmmmrcqntrzlfrljrorjiw