UCL Cancer Institute

Thermo Scientific Open Biosystems pGIPZ shRNAmir library

Individual hairpin clones

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 and can also be titered on your cell type of interest if required. 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 Human 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

£4.00




High quality DNA
miniprep	£25.00
maxiprep	£50.00




Lentiviral particles	unconcentrated (15ml)	concentrated (1ml from 30ml)	concentrated (1ml from 45ml)
with miniprep	£125.00	£175.00	£190.00
with maxiprep	£150.00	£200.00	£215.00




* Viral particles are titrated on 293T cells. Please contact us if you would like your viral particles titered on an alternate cell type also.

** Viral partilces are provided in DMEM (unconcentrated) and Optimem (concentrated). If you would like your virus in alternative media this can be arranged.

*** Please contact us directly for any further requests.



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 GIPZ hairpins within our collection and all Mouse GIPZ hairpins that have Clone IDs starting with V2LHS or V3LHS.
• 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 or V3LHS.
• 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.

Scientific Support Services has a drop-off for sequencing within the Cancer Institute and ICH and will sequence the hairpins for £6 each. You simply need to submit the DNA and indicate that it is a hairpin and requires the GIPZ primer. For more information please refer to the DNA sequencing website

If you wish to sequence your own DNA, you should use the primer:


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