The vectors for Ter HR reporters described were constructed by conventional cloning methods using a previously described 6xTer-HR and RFP-SCR reporters10 (link),26 (link). pHIV-NAT-CD52 vectors were derived from pHIV-Zsgreen, a gift from Bryan Welm and Zena Werb (Addgene plasmid # 18121) 52 (link). Ter-containing plasmids were amplified in JJC33 (Tus–) strains of E. coli. siRNA SMARTpools were purchased from Dharmacon. All plasmids used for transfection were prepared by endotoxin-free maxiprep (QIAGEN Sciences, Maryland, MD).
Protocol detail
Constructing Ter HR Reporter Plasmids
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Beth Israel Deaconess Medical Center, Howard Hughes Medical Institute, Harvard University, Boston Children's Hospital, Jackson Laboratory, The University of Texas Health Science Center at San Antonio
Top 5 similar protocols
Variable analysis
independent variables
- Construction of vectors for Ter HR reporters using conventional cloning methods
- Derivation of pHIV-NAT-CD52 vectors from pHIV-Zsgreen
- Amplification of Ter-containing plasmids in JJC33 (Tus–) strains of E. coli
- Use of siRNA SMARTpools purchased from Dharmacon
- Not explicitly mentioned
- Use of a previously described 6xTer-HR and RFP-SCR reporters as the basis for constructing the Ter HR reporters
- Use of pHIV-Zsgreen, a gift from Bryan Welm and Zena Werb, as the basis for deriving the pHIV-NAT-CD52 vectors
- Preparation of all plasmids used for transfection by endotoxin-free maxiprep (QIAGEN Sciences, Maryland, MD)
- Not explicitly mentioned
- 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.
Sign up for free or login to display all annotations
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!