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Showing: 1 - 14 of 14 results

  1. Lentivirus ddPCR Titration

    Type
    Protocol
    ...aspirate supernatant. Extract genomic DNA according to the GeneJet Genomic DNA Purification Kit instructions...cellular genome: Since the cellular gene RPP30 is used as a reference to the cellular genome, it is assumed... Thermo Scientific, 10199-452 Reagents GeneJet Genomic DNA Purification Kit, Thermo Fisher, K0721 6-well...copies\ RRE \over copies\ RPP30}$$ Use the viruses/genome to calculate the infectious titer: $$T = {V*C*D...Where: T = Infectious titer, TU/mL V = Viruses per genome C = # Cells per well D = Dilution factor v = Virus...serially diluted and used to transduce HEK293T cells. Genomic DNA is extracted from the target cells and assayed...20 µL RRE (FAM) Copies/20 µL RPP30 (HEX) Viruses/genome Infectious titer (TU/mL) Average 1 800 428 22020...

  2. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...brings the pH back down causing the proteins and genomic DNA to precipitate, while leaving the smaller plasmids...plasmids free in solution. Pellet the proteins and genomic DNA by centrifugation, and remove the plasmid-containing... purified away from the bacterial proteins and genomic DNA. Depending on the method used, the DNA concentration...denatured. Pro-Tip Do not vortex at this stage or the genomic DNA will become sheared and will therefore contaminate...formed which contains the bacterial proteins and genomic DNA. Incubate tube on ice for 5 min. Centrifuge...

  3. Video Library

    Type
    Protocol
    ...Storing and Handling Plasmids Genomic Deletions with CRISPR Generate genomic deletions in mammalian cells...cells lines using CRISPR/Cas9 Genomic Deletions with CRISPR Protocol Over-Agar Antibiotic Plating Create...

  4. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Determine the number of genome-containing particles in an AAV prep using qPCR, SYBR Green Technology,...protocol can be used to determine the number of genome-containing particles of an AAV prep using SYBR ...Determine the physical titer of samples (viral genomes (vg)/mL) based on the standard curve and the sample...

  5. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...while others have >1 copy of the transgene in their genome. This can result in varying expression levels of...#52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE...

  7. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...empty capsids. The 60% step serves as a cushion for genome-containing virions. Phenol red is added to clearly...This method will help enrich the prep with full (genome containing) particles. On average, iodixanol-purified...

  8. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... amplifying from plasmid DNA (rather than from genomic DNA or a cDNA library), roughly 18-21bp is usually...increase the Tm if you are trying to clone from genomic DNA, a cDNA library, or by RT-PCR. Isolate your...

  10. Gibson Assembly Protocol

    Type
    Protocol
    .... Chemical synthesis of the mouse mitochondrial genome. Nature Methods , 7(11), 901–903. https://doi.org...

  12. AAV ddPCR Titration

    Type
    Protocol
    ...AAV titer range of 5E+12–5E+13 GC/mL, where GC is genome copies and generally represents the physical number...

  13. CRISPR Library Amplification

    Type
    Protocol
    ...RNAs targeting different genes in an organism's genome, for example. Amplification is usually necessary...

  14. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... DNA is unlikely to be integrated into the host genome. For stable loss-of-function experiments, Addgene... The RNAi Consortium (TRC): www.broad.mit.edu/genome_bio/trc/rnai.html Background on RNAi mechanism: RNAi...
Showing: 1 - 14 of 14 results