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pLVTHM
(Plasmid #12247)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 12247 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pLVTH
  • Backbone size (bp) 11087
  • Vector type
    Mammalian Expression, Lentiviral, RNAi, Cre/Lox

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Stbl3
  • Growth instructions
    Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    None

Cloning Information

Resource Information

Depositor Comments

pLVTHM allows for direct cloning of shRNAs into the lentiviral vectors and replaces the old pLVTH system from Trono lab.
pLVTHM is similar to pLVTH, but contains a 3bp substitution that generates a unique MluI site for direct cloning of an shRNA into MluI-ClaI.

Please note that there is an additional ClaI site in this vector that is blocked by Dam methylation (this site does not appear in the depositor's full sequence). The plasmid needs to be grown in a Dam+ bacterial strain in order to use ClaI for cloning.

Note that ClaI has lower salt concentration requirements than MluI. One way to handle this is to digest with ClaI for 45 minutes, followed by addition of diluted MluI buffer and MluI, then incubation for 90 more minutes. Be sure not to use a large amount of vector (1.7 ug to 2.5 ug is known to work).

Also, if your shRNA is already in pSUPER (or another plasmid under the control of the PolIII promoter) you may use the EcoRI-ClaI sites to replace the H1 promoter in pLVTHM with the H1-shRNA cassette from pSUPER (or another plasmid).

The packaging plasmid for Trono lab lentiviral vectors is also available at Addgene http://www.addgene.org/rnaitools

Please note that the full sequence for this plasmid is approximated and not fully verified. Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more. See LentiWeb http://www.lentiweb.com for discussions on cloning strategies and protocols.

How to cite this plasmid ( Back to top)

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:

    pLVTHM was a gift from Didier Trono (Addgene plasmid # 12247)
  • For your References section:

    Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference. Wiznerowicz M, Trono D. J Virol. 2003 Aug . 77(16):8957-61. 10.1128/JVI.77.16.8957-8951.2003 PubMed 12885912