...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...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning vector. The vector is 8901 base...Protocols pLKO.1 Protocol pLKO.1 - TRC Cloning Vector Addgene Plasmid 10878. Protocol Version 1.0. December 2006...pLKO.1-TRC Cloning Vector A.1 The RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA...

...that have taken up the lentiviral transgene. Note that not all lentiviral vectors deliver antibiotic resistance...an integrated lentiviral vector. Protocols...expressing a gene of interest from an integrated lentiviral vector. Unlike the short-term protein expression ...transfection approaches, generating cell lines using lentiviral vectors enables long-term protein expression studies...with repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic resistance (...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries...Stable Cell Lines with Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols...

...from a lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol... 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume 500 μL *Plasmid concentrations and ... each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid DNA purification...endotoxin removal step. For high quality plasmid DNA, the plasmids should also be propagated in an endonuclease... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...Viral Vector Guides Mol Bio Protocols Introduction This protocol can be used to produce lentivirus from...~20 h. Prepare a mixture of the 3 transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish...

... of Lentivirus Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence... (ddPCR) for lentivirus vector titration. Protocols... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration... use droplet digital PCR (ddPCR) to titer lentivirus vectors. This protocol specifically uses primers ...ddPCR Lentivirus sample data, RPP30 positive (green) and negative (black) droplets Sample ID Plasmid ID ...be close to zero (A01). In this protocol, the lentiviral particles are serially diluted and used to transduce...Y, Bergelson S, Feschenko M. Determination of Lentiviral Infectious Titer by a Novel Droplet Digital PCR...

...complicated procedures. Plasmid Cloning Protocols for constructing and analyzing your plasmids. Virus Protocols...Amplify CRISPR pooled-plasmid libraries Diagnostic Restriction Digest Verify a plasmid using restriction ...polymerase pLKO.1 - TRC Cloning Vector Cloning protocols for using the pLKO.1 vector, a backbone used by the ...Transfection Introduce plasmid DNA to mammalian cells Lentivirus Production Produce lentivirus with a polyethyenimine... that can be used for basic molecular biology, plasmid cloning, and titering and testing your viral preparations...Bacterial Glycerol Stocks Store bacterial strains or plasmids for long term use Watch the Video! DNA Purification...spectrophotometer Restriction Digests Cut and analyze plasmid DNA with restriction enzymes Agarose Gel Electrophoresis...

...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries...by Limiting Dilution You may also like... Viral Vector Guides Virus Blog Posts Addgene Protocols Viral... pool of stable cells. Transducing cells with lentivirus results in a heterogenous polyclonal population...protocol for generating stable cell lines with lentivirus . Day 0: (optional) Seed cells for the generation...protocol for generating stable cell lines with lentivirus . Pro-Tip Because the polyclonal cell pool is...