Generation of AR and AR-V7 Lentiviral Constructs

DH5a and TOP10 competent cells were purchased from Thermo Fisher Scientific, Waltham, MD and used for plasmid transformation and propagation as per manufacturer’s instructions. PCR amplified cDNA of AR (full length) was cloned from an available pCR3.1 expression vector^{50 (link)} by fusing with 3X FLAG sequences at the beginning and an HA-Tag at the end. This was cloned into the pHAGE lentiviral expression vector as previously described^{25 (link)}. pCR3.1 AR-V7 has been described previously^{51 (link)}. The pLX302_FOXA1-V5 plasmid was purchased from Addgene, Watertown, MA.

Free full text: Click here

Basil P., Robertson M.J., Bingman WE I.I.I., Dash A.K., Krause W.C., Shafi A.A., Piyarathna B., Coarfa C, & Weigel N.L. (2022). Cistrome and transcriptome analysis identifies unique androgen receptor (AR) and AR-V7 splice variant chromatin binding and transcriptional activities. Scientific Reports, 12, 5351.

Publication 2022

TOP10 competent cells

Cdna Cells Foxa1 Phage Plasmid Vector

Corresponding Organization : Baylor College of Medicine

Top 5 similar protocols

Variable analysis

independent variables

Transformation of DH5a and TOP10 competent cells with plasmids
Cloning of AR (full length) cDNA from pCR3.1 expression vector and fusing it with 3X FLAG and HA-Tag sequences
Cloning of AR-V7 from pCR3.1 AR-V7 plasmid
Acquisition of pLX302_FOXA1-V5 plasmid from Addgene

dependent variables

Plasmid transformation and propagation

control variables

Manufacturer's instructions for transformation and propagation of DH5a and TOP10 competent cells
Procedures previously described for cloning AR (full length) into pHAGE lentiviral expression vector
Previous description of pCR3.1 AR-V7 plasmid

controls

Positive control: pCR3.1 expression vector containing AR (full length) cDNA
Positive control: pCR3.1 AR-V7 plasmid
Positive control: pLX302_FOXA1-V5 plasmid from Addgene

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!