Purpose3rd gen lentiviral negative control vector containing scrambled shRNA
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||1864||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerStewart SA, RNA 2003 Apr; 9(4):493-501.
- Backbone size w/o insert (bp) 7032
Vector typeMammalian Expression, Lentiviral, RNAi
Growth in Bacteria
Growth Strain(s)XL10 Gold
Growth instructionsXL10-Gold Ultracompetent Cells from Stratagene. 37oC.
Insert Size (bp)60
- Cloning method Restriction Enzyme
- 5′ cloning site AgeI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer LKO.1 5' (Common Sequencing Primers)
shRNA for use as a negative control. Sequence of hairpin is:CCTAAGGTTAAGTCGCCCTCGCTCGAGCGAGGGCGACTTAACCTTAGG. For packaging, please use pCMV-dR8.2 dvpr (Addgene plasmid #8455) and pCMV-VSVG (Addgene plasmid #8454).
Please note that the 5' cloning site, AgeI, is typically destroyed during the shRNA cloning. Depending on the specific shRNA sequence, the site can occasionally be restored. AgeI is present in this plasmid.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:scramble shRNA was a gift from David Sabatini (Addgene plasmid # 1864 ; http://n2t.net/addgene:1864 ; RRID:Addgene_1864)
For your References section:Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Sarbassov DD, Guertin DA, Ali SM, Sabatini DM. Science 2005 Feb 18;307(5712):1098-101. 10.1126/science.1106148 PubMed 15718470