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Showing: 481 - 520 of 731 results
  1. Protocol - Bacterial Transformation

    Type
    Protocol
    ... for Ampicillin resistance but is much more important for other antibiotic resistances. Plate some or ...bacterial origin of replication and an antibiotic resistance gene for use as a selectable marker in bacteria... the bacteria time to generate the antibiotic resistance proteins encoded in the plasmid backbone so that...Shorten or skip the outgrowth (for Ampicillin resistance it is ok to completely skip the outgrowth, for... plate containing the correct antibiotic. The resistance gene on your plasmid must match the antibiotic...
  2. Video Library

    Type
    Protocol
    ... Technology Protocol Agarose Gel Electrophoresis Gel electrophoresis is the standard laboratory procedure... once using a multichannel pipette and avoid inconsistent pipetting with our simple tips Bacterial Transformations...visualization and purification. Agarose Gel Electrophoresis Protocol Purifying DNA from an Agarose Gel ...DNA fragments based on size. Following gel electrophoresis, you can cut DNA bands out of the agarose gel...
  3. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...gel electrophoresis, which separates DNA by their length in base pairs. Following electrophoresis, you...Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis DNA Ligation Background Information Gel purification...Protocol: Gel Purification Follow the Agarose Gel Electrophoresis Protocol with the following amendments: Notes...
  4. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ... carry one or more antibiotic resistance genes, which confer resistance to a specific antibiotic to the... carrying them. The presence of an antibiotic resistance gene on a plasmids allows researchers to easily...antibiotic in your LB media matches the antibiotic resistance on your plasmid. If the bacteria on your LB agar...
  5. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...the Video! Sequence Analysis Verify important plasmid features using sequence analysis RNA Extraction Without...plasmid DNA with restriction enzymes Agarose Gel Electrophoresis Separate DNA by size on an agarose gel Watch...Lentivirus Quantify virus by counting antibiotic resistant colonies Generating Stable Cell Lines with Lentivirus...
  6. Gibson Assembly Protocol

    Type
    Protocol
    ...plasmids by designing primers to split an antibiotic resistance gene to effectively create an extra part, one... Assembly master mix (isothermal reaction mix) consists of three different enzymes within a single buffer...Assembly® is licensed to New England Biolabs by TelesisBio. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison...Hutchison CA, Venter JC, Merryman C. (2010). Chemical synthesis of the mouse mitochondrial genome. Nature Methods...
  7. Colony Formation Titering Assay

    Type
    Protocol
    ...lentiviral preparations that confer antibiotic resistance....lentivirus preparations that confer antibiotic resistance. Determining the titer of your lentiviral preparation...selection markers. Note this assay requires staining resistant colonies with crystal violet solution and, therefore...media as needed Days 14–18: Stain cells and count resistant colonies Equipment Class II, Type A2 Biological...
  8. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min Equipment qPCR instrument Heating plate... #59462 is one plasmid that gave reliable and consistent results. Use the recommended plasmid, or test... Sample 4 F E Sample 2 Sample 5 F Perform data analysis using the instrument’s software. Determine the...determine differences between samples. Melt curve analysis: a single peak should be seen. The presence of...
  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Polyethylene Glycol 8000 (PEG) + 0.5 M NaCl Cell lysis buffer (50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl2)...separate into phases. Aliquot and store at 4 °C. Cell Lysis Buffer : 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl...resuspend the pellets in a total of 5 mL of cell lysis buffer (recipe above). Pipet back and forth to resuspend... the cells by adding a total of 12.5 mL of cell lysis buffer. Pipet back and forth to resuspend each pellet...
  10. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...target cell line. Once introduced, the puromycin resistance marker encoded in pLKO.1 allows for convenient...drives expression of puromycin. Puro R Puromycin resistance gene for selection of pLKO.1 plasmid in mammalian...bacterial origin of replication. Amp R Ampicillin resistance gene for selection of pLKO.1 plasmid in bacterial...allow for sufficient expression of the puromycin resistance gene. f. To select for infected cells, add puromycin...you as a guide and on an “AS IS” “AS AVAILABLE” basis without warranty of any kind either expressed or...
  11. We’re Going Viral This Week!

    Type
    Blog Post
    ...strategies to improve viral vector production and data analysis. Our first paper came out last year on sucrose...
  12. Summer Fun at Addgene!

    Type
    Blog Post
    ...tackling complex questions, and giving tips for consistently generating solid data. If you’re ready for the...
  13. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...containing the carbenicillin (amp) resistance gene (or other antibiotic resistance) Procedure Day 1 Prepare carbenicillin... cells containing plasmids differing in their resistance genes, as one does not need to prepare separate...
  14. A Scientist's Guide to Ebola

    Type
    Blog Post
    ...scientific literature.  -In light of the current Ebola crisis, Science and Science Translational Medicine have...
  15. Let There Be LITE Plasmids

    Type
    Blog Post
    ...ingredients - CRY2 and CIB1 – which they borrowed from Arabidopsis thaliana. The results are light-inducible transcriptional...
Showing: 481 - 520 of 731 results