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...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...

...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...

... 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 plasmid allows researchers to easily...antibiotic in your LB media matches the antibiotic resistance on your plasmid. If the bacteria on your LB agar...

...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...

...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...

...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...

...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...

...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...

...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...

...itself. Many of us have experienced the minor yet consistent annoyance of walking into a fly lab and finding...

...strategies to improve viral vector production and data analysis. Our first paper came out last year on sucrose...

...published their work on a protein required for cytokinesis in JEI. Developing an interested in biology, ...

... first on a temporary and now on a more lasting basis.   Table 1: Popular Rabies and VSV-G Plasmids ...

...Resources on Addgene.org Use the Kit-Free Alkaline Lysis Plasmid Miniprep Protocol Learn How to Analyze Your...

...While not technically a research story, I couldn't resist adding in this excellent Q&A from Tim Miles on ...

...pieces of energy-intensive lab equipment on a daily basis. By the beginning of September all will be powered...

...make a big impact in open science. Depositors consistently contribute to our collection so that we can ...

...tackling complex questions, and giving tips for consistently generating solid data. If you’re ready for the...

...like... Bacterial Transformation Agarose Gel Electrophoresis Agarose Gel DNA Purification Streaking and ...purity click here . Top Protocol: Kit-free Alkaline Lysis Plasmid Miniprep Prepare the following solutions...phase Middle phase - A white layer may appear, consisting of precipitated protein particles Bottom phase...

...metrics!  About the Author Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...