UCL Cancer Institute

Open Biosystems pGIPZ shRNAmir library


Individual hairpin clones


These GIPZ hairpins and any derivatives (viruses, cell lines, portions of the GIPZ vector cloned into other vectors etc.) must not be shared or transferred to Institutes outside UCL. They must remain and be used solely within UCL laboratories

    Formats


    Individual hairpins come as either bacterial agar stab cultures, high quality DNA ready for transfection or lentiviral particles ready for transduction.

    • Bacterial stab cultures: You will need to re-streak the bacteria from the surface of the culture onto an LB-Amp plate, pick a single colony, grow, prepare and purify your plasmid DNA using a kit and sequence the DNA to make sure the sequence is correct. The DNA is then ready for transfection.

    • High quality DNA: Your hairpins will come as a high quality DNA mini or maxi-prep that has been sequence confirmed and is ready for transfection. You will also be provided with a glycerol stock, sequence comparison and quality metrics of your DNA prep.

    • Lentiviral particles: Your hairpins will come as lentiviral particles in culture media, either concentrated or unconconcentrated depending on your requirements, that is ready for transduction of your cell type of interest. This will have been titered on 293T. You will also be provided with the remaining high quality DNA mini or maxi-prep, a glycerol stock, sequence comparison and quality metrics of your DNA prep.

    Controls


    We have two negative controls and two positive controls for the GIPZ library. Please see the FAQs section for additional sequence details of these controls.

    • Negative:
      • Empty GIPZ vector
      • Non-silencing (scrambled) (22mer: aTCTCGCTTGGGCGAGAGTAAG)
    • Positive
      • GAPDH (22mer: cCCTCATTTCCTGGTATGACAA)
      • EG5 (22mer: cGGCCATGCTAGAAGTACATAA) Targets Human but not Mouse

    Controls are the same price as the shRNA hairpins and can be ordered using the order form below

    UCL Consortium prices


    This includes members of the UCL Cancer Institute, Institute of Child Health, Institute of Neurology and the Division of Infection and Immunity.

     

    Bacterial stab cultures £15.00

    High quality DNA  
    miniprep £55.00
    maxiprep £85.00

    Lentiviral particles unconcentrated (15ml) concentrated (1ml from 30ml) concentrated (1ml from 60ml)
    with miniprep £195.00 £240.00 £265.00
    with maxiprep £225.00 £270.00 £295.00

    *If you would prefer an alternative volume of virus produced to what is outlined above, please contact us for a quote.

    *All viral particles are provided in DMEM (unconcentrated) and Optimem (concentrated) and are tittered on 293T cells. If you would like your virus produced in an alternative media or tittered on an alternative cell type this can be arranged, please just contact us for further details.

    Ordering clones

     

    Instructions:

    • Download the GIPZ Order Form
    • Go to the Thermo Scientific website and follow these GIPZ Ordering Instructions to search for available hairpins. We have all of the Human and Mouse GIPZ hairpins within our collection starting with the codes V2LHS, V3LHS, V2LMM or V3LMM.
    • Take note of the CloneID for the hairpins you would like
    • If you are not from within the Cancer Institute, please arrange an eIDT assigned to Catherine King at the Cancer Institute to cover the cost of the order. Refer to the price list above or if you are unsure please contact us
    • Complete the ordering form:
        • Make sure you enter your grant code (CI users) or eIDT number (all other users)
        • Indicate whether you would like any controls and what format you would like
        • List the the CloneID of the hairpins you are after. These codes will start with V2LHS, V3LHS, V2LMM or V3LMM.
    • Email your completed form to rnai@cancer.ucl.ac.uk.
    • We will contact you to let you know your order has been received and to arrange collection. All clones are collected from the UCL Cancer Institute, Paul O'Gorman Building on Huntley St.

    • Sequencing your hairpins


      We highly recommend sequencing your hairpins before starting work with them.

    • GIPZ sequencing primer: 5' - GCATTAAAGCAGCGTATC - 3' Melting Temp: 52.7c The binding site for this primer is base 5820-5842 and runs in reverse complement direction.

 

Contacts


Catherine King
0207-679-6506
catherine.king@ucl.ac.uk

 



 

Links


Please refer to the Thermo Scientific website for more detailed information about our library.