shRNA

Technical Bulletins

Bacterial Glycerol Stock (SHCLNG)
Purified Plasmid DNA Format (SHCLND)
Lentiviral Format (SHCLNV)
MISSION® Control Vectors
MISSION Control Transduction Particles
MISSION Lentiviral Packaging Mix (SHP001)
MISSION 3'UTR Lenti GoClone™

 

Bacterial Glycerol Stock (SHCLNG)
Bacterial cultures may be amplified from the glycerol stocks for use in purification of shRNA plasmid DNA. Subsequently, target cell lines may be transfected with the purified plasmid for transient or stable gene silencing (puromycin selection). In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids

Individual shRNA clones are provided as a frozen bacterial glycerol stock (E. coli) containing Terrific Broth (TB), carbenicillin at 100 µg/ml and 10% glycerol.

Download the SHCLNG Technical Bulletin (583 kb PDF)

 


Purified Plasmid DNA Format (SHCLND)
Sigma offers the highest quality purified plasmid DNA for downstream transfection and lentiviral packaging. DNA purified with Sigma’s 96-well DNA purification system, developed specifically for the TRC library, yields the highest quality lentiviral particles.

Purified plasmid may be directly used in transient or stable gene silencing (puromycin selection). In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids.

Individual constructs are provided as frozen stocks containing 1 µg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 20 ng/µL. See gene family set technical bulletins for specifications, inquire for library specifications.

Download the SHCLND Technical Bulletin (561 kb PDF)

 


Lentiviral Format (SHCLNV)
Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids. In addition, the lentiviral transduction particles are pseudotyped with an envelope G glycoprotein from Vesicular Stomatitis Virus (VSV-G), allowing transduction of a wide variety of mammalian cells. The lentiviral transduction particles are titered via a p24 antigen ELISA assay and pg/ml of p24 are then converted to transducing units per ml using a conversion factor.

Individual constructs are provided as a 200 µL frozen stock containing a minimum of 1.0 x 106 TU/ml (via p24 assay) in Dulbecco’s Modified Eagle’s Medium with 10% heat-inactivated fetal bovine serum and penicillin-streptomycin. Sigma’s average titers for individual clone orders are routinely greater than 1.0 x 107 TU/ml (via p24 assay). See gene family set technical bulletins for specifications, inquire for library specifications.

Download the SHCLNV Technical Bulletin (124 kb PDF)

 


MISSION Control Vectors

Purified plasmid control vectors may be directly used in transient or stable gene silencing (puromycin selection). In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids. Visit our controls webpage for more information on specific controls.

Individual control vectors are provided as 10 µg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/µl.

Download the MISSION Control Vectors Technical Bulletin (226 Kb PDF)

 


MISSION Control Transduction Particles

Sigma offers a variety of positive, negative, and transduction controls in lentiviral format. Additionally, our most popular controls are offered in high titer format. Visit our controls webpage for more information on specific controls.

Standard titer control transduction particles are provided as 200 µl of 106 TU/ml (via p24 titering assay). High titer control transduction particles are provided as 10 x 20 µl aliquots of 109 TU/ml (via p24 titering assay).

Download the MISSION Control Transduction Particles Technical Bulletin (231 Kb PDF)

 


MISSION Lentiviral Packaging Mix (SHP001)

The MISSION Lentiviral Packaging Mix is an optimized formulation of two plasmids expressing the key HIV packaging genes and a heterologous viral envelope gene. It is designed to be co-transfected along with a compatible lentiviral transfer vector in order to create high-titer pseudo-typed lentiviral particles used for downstream transduction applications.

The MISSION Lentiviral Packaging Mix is available in two package sizes: SHP001-0.25 mL and SHP001-1.7 mL.

Download the MISSION Lentiviral Packaging Mix Technical Bulletin (286 Kb PDF)

 


MISSION 3'UTR Lenti GoClone

The 3' untranslated regions (3'UTRs) of mRNA transcripts control many aspects of post-transcriptional regulation of genes. 3'UTRs are involved in regulating transcript stability, translational efficiency, and localization in the cell. 3'UTRs are also the predominant target sequences for endogenous microRNAs (miRNA) or small interfering RNAs (siRNA).

Sigma-Aldrich and SwitchGear Genomics have teamed up to offer you the MISSION 3'UTR Lenti GoClone collection, which is a genome-wide collection of cloned human 3'UTRs. Each viral vector expresses an optimized luciferase reporter gene (RenSP) fused to a 3’UTR sequence.

Download the MISSION 3'UTR Lenti GoClone Technical Bulletin (154 Kb PDF)

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