Lentiviral Particle Production and Titration

Lentiviral plasmids were cotransfected into HEK293T cells using calcium phosphate precipitation as previously described [3 (link)–5 (link)]. Lentiviral particles were harvested at 36, 48, 60, and 72 h posttransfection, filtered through Millex-HV 0.45 μM polyvinylidene fluoride syringe filters (EMD Millipore^©, Burlington, USA), and concentrated using Amicon Ultra 100 kDa filters (EMD Millipore^©, USA). Concentrated lentiviral particles were titered using NIH3T3 cells and FACS analysis of eGFP and mCherry expression. Flow cytometry data were analyzed using BD FACSDiva™ software (Version 8.0.1).

Free full text: Click here

Gerace D., Martiniello-Wilks R., Habib R., Ren B., Nassif N.T., O'Brien B.A, & Simpson A.M. (2019). Ex Vivo Expansion of Murine MSC Impairs Transcription Factor-Induced Differentiation into Pancreatic β-Cells. Stem Cells International, 2019, 1395301.

Publication 2019

Amicon Ultra 100 kDa filters BD FACSDiva™ software

Calcium phosphate Cells Flow cytometry Nih3t3 cells Plasmids Polyvinylidene fluoride Syringe

Corresponding Organization : University of Technology Sydney

Top 5 similar protocols

Variable analysis

independent variables

Lentiviral plasmids

dependent variables

Lentiviral particle titer
EGFP and mCherry expression

control variables

Calcium phosphate precipitation protocol
Millex-HV 0.45 μM polyvinylidene fluoride syringe filters
Amicon Ultra 100 kDa filters
NIH3T3 cells
BD FACSDiva™ software (Version 8.0.1)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!