We narrowed to 7 results for: Cre expression vector
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TypeProtocol...stable cell lines expressing a gene of interest from an integrated lentiviral vector....stable cell lines expressing a gene of interest from an integrated lentiviral vector. Unlike the short-term...using lentiviral vectors enables long-term protein expression studies. Moreover, repeating experiments ... the most desirable level of expression. Over time, transgene expression in a polyclonal population may...is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, ...Lines with Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...short-term protein expression observed using transient transfection approaches, generating cell lines using...
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Plasmid Cloning by Restriction Enzyme Digest (with Protocols)
TypeProtocol...bacterial expression vector. Using subcloning, you can easily move YGOI into a mammalian expression vector...same orientation on your target vector. If you are not sure what vector to use, you can check out our Empty...Congratulations, you now have YGOI in a mammalian expression vector and can begin your studies. Reference Page...be used to easily move any piece of DNA from one vector to another as long as it is already bounded by ...of interest (YGOI for short). You might need to express YGOI in cultured mammalian cells. The problem is...orientation in the recipient plasmid. (You don't want to express the antisense version of your gene!) Are in frame...manufacturer’s instructions. Isolate your insert and vector by gel purification Run your digested DNA on an... -
Protocol - pLKO.1 – TRC Cloning Vector
TypeProtocol...TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning vector. The vector is 8901 base...Cloning Vector The following steps were the prevailing guidelines when the pLKO.1 vector was created. For...-Cloning Vector I.2 Recipes I.3 Warranty information Back to Top A. pLKO.1-TRC Cloning Vector A.1 The ...use of this vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector chosen by the...TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector containing... pLKO derived vectors that may also be useful for your experiment. To see these vectors, visit Addgene...and ligated into the pLKO.1 vector, producing a final plasmid that expresses the shRNA of interest. C.1... -
Isolating a Monoclonal Cell Population by Limiting Dilution
TypeProtocol...could lead to reduced transgene expression over time, as the lower expressing clones take over the polyclonal...have been sufficiently expanded, screen the lines for transgene expression and/or other phenotypes. For example...Western blotting to screen for lines with the highest or lowest transgene expression ( Figure 2 ). Sample...is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, ...by Limiting Dilution You may also like... Viral Vector Guides Virus Blog Posts Addgene Protocols Viral...that are more likely to retain stable transgene expression. Other methods of generating a monoclonal cell...population be monitored regularly for transgene expression. This protocol has been optimized for adherent... -
Fluorescence Titering Assay
TypeProtocol...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...marker. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection...protocol was developed using low titer lentiviral vectors (10 5 TU/mL). If you anticipate that your viral... transduction, cells were assayed for mCherry expression using fluorescence microscopy. Last reviewed ... remove cells and debris. Lentiviral titer can decrease during cycles of freeze-thaw. If you are freezing... -
Protocol - Bacterial Transformation
TypeProtocol...plasmids (even those designed for mammalian cell expression) carry both a bacterial origin of replication...approximately 10-fold lower for ligation of inserts to vectors than for an intact control plasmid. Incubate the...Scientists have made many genetic modifications to create bacterial strains that can be more easily transformed... specific treatments have been discovered that increase the transformation efficiency and make bacteria... -
AAV Production in HEK293 Cells
TypeProtocol...Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference...transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene of interest Triton X-100 Benzonase/...harvesting. The addition of sorbitol has been shown to increase viral titers by up to 1.8-fold ( Rego et al., ...sodium bicarbonate, sorbitol has been found to increase viral titers by up to 1.8-fold. See the recipe...