pRetro-U6G vector, please submit your gene name, accession #, target and
loop sequences here.
conditions listed on the ordering page. Submitting an order form is construed as
you have carefully read and agree with such terms and conditions.
Purchase of the shRNA vector Construction service amounts to entering a service
agreement under which Cellogenetics is contractually obligated to:
1. synthesize the corresponding DNA oligoes for and generate the ordered shRNA
expression constructs.
2. confirm the cloned shRNA coding sequence by DNA sequencing.
3. make glycerol stocks of bacterial clones containing the ordered shRNA
expression constructs and prepare technical spec sheet.
4. ship the ordered shRNA expression constructs as glycerol stocks along with all
pertinent technical information within 15 business days from the date when the
order was placed.
modified from murine embryonic stem cell virus (pMSCV)
plasmid to express double stranded small hairpin RNA
from human U6 snRNA promoter. A compounded
puromycin resistance-internal ribosome entry site-green
fluorescence protein (puro-IRES-GFP) cassette is inserted
behind the phosphoglycerate kinase (PKG) promoter for
antibiotic selection and green fluorescence labeling in
mammalian cells. Annealed DNA oligos for expression of
small hairpin RNAs are to be cloned behind the human
U6 snRNA promoter between BamHI and Xho I sites. The
small hairpin RNA encoded by pRetro-U6G can be
delivered either by standard plasmid transfection
approach or by infecting target cells with packaged virus
particles. The replication incompetent retroviral particles
generated upon transfection of packaging cells, such as
Phoenix A or E cells, can infect a wide range of target
cells, including hematopoietic and embryonic stem cells,
as well as most other mitotically active cells, and
achieves high level expression of small hairpin RNAs.
All pRetro-U6G derived plasmids should be propagated in
suitable host strains such as DH5a, or HB101, under
selection of resistance to ampicillin (50 mg/ml). E.coli
replication origin: pUC; copy number: high.
