Retroviral Lentiviral transduction protocolStewart Lab
Millipore, Billerica, MA)between day 18 and 20 posttransfection. One of the fastest and most reliable methods for selecting a reprogrammed colony is live staining withTra1-60 or Tra1-81 antibodies.
pLKO.1 can be introduced into cells via direct transfection, or can be converted into lentiviral particles for subsequent infection of a target cell line. Once introduced, the puromycin resistance marker encoded in pLKO.1 allows for convenient stable selection. Figure 1 : Map of pLKO.1 ...
Lentiviral infection 1. Day 4 (~ 4pm): For fibroblast cells, replace with 2ml fresh medium 2h in advance of transduction. 2. Day 4 (~ 6pm) Add equal volume (800ul) supernatant of each factor ( Yu1, Yu2, Yu3) to fibroblast dish in the presence of 10ug/ml polybrene, incubate at ...
materials D.2 Screening for inserts E. Producing Lentiviral Particles E.1 Recommended materials E.2 Protocol for producing lentiviral particles F. Infecting Target Cells F.1 Recommended materials F.2 Determining the optimal puromycin concentration F.3 Protocol for lentiviral infection and selection G....
Development of optimal bicistronic lentiviral vectors facilitates high-level TCR gene expression and robust tumor cell recognition. Gene Ther. 15 (21), 1411-1423 (2008). Froger, A., Hall, J. E. Transformation of plasmid DNA into E. coli using the heat shock method. J. Vis. Exp. (6),...
1 x önemli olmayan amino asit, 2 mM L-glutamin) % 5 ile 37 ° C'de CO2. protokol tekrarlanabilirlik için bu farklı bir çok/toplu işlemine geçiş yaparken buzağı serum test etmek için tavsiye edilir. Altı 15 cm kadar tabak lentiviral üretim için ihtiya...
(Cat. Nos. T100A, T100B & T202) for high-efficiency retroviral or lentiviral transduction of hematopoietic cells. After transduction, the cells can then be expanded into the largerGT-T610 (CultiLife Eva) bag(Cat. # FU0010), which is composed of ethylene vinyl acetate (EVA), a flexible ...
F.InfectingTargetCellsoF.1RecommendedmaterialsoF.2DeterminingtheoptimalpuromycinconcentrationoF.3Protocolforlentiviralinfectionandselection G.Safety---.pLKOshRNA病毒质粒载体构建protocol方法详解 H.ReferencesoH.1PublishedarticlesoH.2Webresources I.AppendixoI.1SequenceofpLKO.1TRC-CloningVectoroI.2RecipesoI.3Warranty...
filtered (0.45 μM), and applied to iHPCs (500 uL/0.25 × 106cells) in the 2.9 medium. Media of iHPCs was fully replaced the next day and the cells were differentiated for 28–42 days. Transduction efficiency was assessed by eGFP fluorescence using an Attune™ Nxt Flow Cytometer...