Positive and negative controls are a critical part of a gene silencing experiment. They enable accurate interpretation of knockdown data and provide confidence in the specificity of the response. We have developed positive and negative controls for the TRIPZ products. These are available as glycerol stock or viral particle formats.
Changes in the mRNA or protein levels in cells treated with negative or non-silencing controls reflect non-specific responses in cells and can be used as a baseline against which specific knockdown can be measured.
Positive controls confirm that the TRIPZ shRNA construct is successfully entering the cell, can be induced by doxycycline and can activate the RNAi pathway.
Available TRIPZ shRNA controls include:
- TRIPZ Inducible Lentiviral Negative shRNA Control: A non-targeting TRIPZ shRNA designed with minimal homology to known mammalian genes.
- TRIPZ Inducible Lentiviral Positive shRNA Control: A TRIPZ shRNA targeting human GAPDH for knockdown and subsequent GAPDH mRNA quantification.
Glycerol stocks of shRNA containing control vectors are shipped as bacterial cultures of E. coli
in LB (low salt) broth with 8% glycerol, carbenicillin (100 µg/mL) and zeocin (25 µg/mL). We check all cultures for growth prior to shipment.
pTRIPZ lentiviral shRNA vector
- Editorial, Whither RNAi? Nature Cell Biology. 5(6), 489-490 (2003).