FAQ

General

How are the prices calculated?
How can I request a quote? How can ask for a order?
Where can I find FuGU Libraries and your personnel?
How can I contact FuGU Libraries?
What is the approximate delivery schedule for different products?
Do I need to have a GMO license for using lentiviral particles in my experiments?

 

Virus Services

Which packaging system is used for lentiviral particle production?
How much plasmid vector DNA is required for viral particle production?
How are the products delivered to me? 
How can I order glycerol stocks from FuGU Libraries?
How should I grow glycerol stocks from TRC / KO gRNA libraries?

shRNA and gRNA Libraries

I would like to order TRC1 library shRNA constructs targeting genes X and Y. Are they available?
I would like to order Sanger KO library gRNA constructs targeting genes X and Y. Are they available?
What kind of selection do the shRNA constructs have?
What kind of selection markers and tags do the gRNA constructs have?
Does FuGU Libraries have different controls for mouse and human TRC lentiviral shRNA libraries?
Which controls do you have for lentiviral gRNA KO libraries?
 

Quality and titer

What is a p24 test?
What is the difference between pg/ml and TU/ml?
What is a RCV test?
How should I store viral particles ordered from FuGU Libraries?
 

General

How are the prices calculated?

FuGU is a non-profit unit of the Faculty of Medicine in the University of Helsinki and the prices come from reagents, labor and equipment costs. The guidelines of pricing are given by the University of Helsinki finances. The faculty of Medicine approves the price list.

How can I request a quote? How can ask for a order?

Please, fill out our Online Order Form or place your request to us by email: fugu-support(at)helsinki.fi
The information we ask from you is described in more detail here.

Where can I find FuGU Libraries and your personnel?

FuGU Libraries is located in Biomedicum 1. Take the main elevators to the 5th floor and turn left to B wing, FuGU Libraries is located in room B508.

How can I contact FuGU Libraries?

FuGU Libraries has a support email address where you can send your request from any service area. Please contact fugu-support(at)helsinki.fi Telephone number: + 358 2 941 25494

What is the approximate delivery schedule for different products?

Glycerol stocks: 2 days - 2 weeks
DNA preparations: 2 days -3 weeks
Lenti/Retroviral particles: 2 - 5 weeks
Concentrated lentiviral particles: 2 - 5 weeks
Q-RT-PCR knockdown/gene expression validation: 2 - 4 weeks

NB! The exact delivery schedule is defined separately for each order.

Does every group need to have their own GMO license? / Do I need to have a license for using lentiviral particles in my experiments?

Yes. Every group/institute is responsible to have its own GMO license. For bacterial glycerol stocks and DNA you have to have a license for biosafety level 1 material and for lenti- and retroviruses you need to have the license for biosafety level 2.

 

Virus services

 

Which packaging system is used for lentiviral particle production?

FuGU Libraries uses second generation lentiviral packaging system. Thus, our packaging system is suitable for both 2nd and 3rd generation transfer vectors.
TRC shRNA library constructs are in 3rd generation transfer vectors.

How much plasmid vector DNA is required for viral particle production?

Following amount of customer DNA is required per one construct:
Mini scale: 5 μg
Midi scale: 10 μg
Concentrated virus: 30 μg (for 180 μl), 60 μg (for 360 μl)

My research group is located in Biomedicum1/Meilahti campus (1.), Viikki (2.), University of Oulu (3.).
How are the products delivered to me?

1. You are welcome to pick-up your order from FuGU Libraries.
2. FuGU Libraries will deliver your products via University of Helsinki internal mail. The products are shipped at noon and should be delivered during the same working day.
3. Your order will be delivered via transportation company, such as DHL, FedEx and TNT. The shipment fee is charged from the customer.

How can I order glycerol stocks from FuGU Libraries?

All customers from Finnish universities can order TRC1 library shRNA clones from FuGU Libraries as bacterial glycerol stocks. Please, fill out our Virus Particle Order form.
FuGU Libraries has Broad Institute's TRC1 lentiviral shRNA library (both human and mouse) in pLKO.1 vector backbone (Broad Institute's TRC library database).

How should I grow glycerol stocks from TRC / KO gRNA libraries?

TRC clones are low copy number plasmids and should be grown in Terrific Broth (TB) including carbenicillin. Clones from Sanger KO gRNA library grow well in LB medium with added ampicillin. We recommend LB medium with ampicillin to produce DNA maxipreparations from clones of both libraries.

 

shRNA and gRNA Libraries

 

I would like to order TRC1 library shRNA constructs targeting genes X and Y. Are they available?

shRNA constructs can be browsed by individual genes here. Contact us, if the construct of interest is part of TRC2 library. When you are making an shRNA construct inquiry/order, please specify whether you are interested in DNA preparations or glycerol stocks and include the Official Gene Symbol (e.g. Entrez gene ID).

I would like to or­der Sanger KO library gRNA con­structs tar­get­ing genes X and Y. Are they avail­able?

KO library gRNA constructs can be browsed by individual genes here. When you are making a gRNA construct inquiry/order, please specify whether you are interested in DNA preparations or glycerol stocks and include the Official Gene Symbol (e.g. Entrez gene ID).

What kind of selection do the shRNA constructs have?

The constructs are in pLKO.1 vector with puromycin selection. Carbenicillin selection is used for the growth and maintenance of the vector in bacterial cells.

What kind of se­lec­tion mark­ers and tags do the gRNA con­structs have?

gRNA construct are in LV04 vector that includes pyromycin selection cassette and a blue fluorescent protein (BFP) tag for positive-cell visualization. Full vector map and sequence can be found here.

Does FuGU Libraries have different controls for mouse and human TRC lentiviral shRNA libraries?

No. Same controls can be used for both mouse and human TRC shRNA libraries. The controls available for TRC library shRNAs are:
Empty pLKO.1 vector; Scramble shRNA in pLKO.1 backbone; TurboGFP in pLKO.1 backbone; eGFP shRNA in pLKO.1 backbone

Please have a look here for further descriptions of above listed controls.

Which con­trols do you have for len­tiviral gRNA KO lib­rar­ies?

We can provide non-targeting gRNA constructs in LV04 vector either with or without Cas9 expression. Same controls can be used for mouse and human KO gRNA libraries. A positive control gRNA construct is also available. Please see the details here.

 

Quality and titer

 

What is p24 test?

The p24 test measures the p24 capsid protein from lentiviruses and gives you a titer pg/ml. p24 test is always made for our internal lentivirus control, for MIDI scale lentiviruses and for concentrated lentiviruses both before and after concentration. Customers can also order the test for viruses they have produced by themselves. The service is provided by BVC unit.

What is the difference between pg/ml and TU/ml?

TU/ml (Transduction units/ml) varies between different cell lines and needs to be determined at each case. It tells you how much virus actually infects a target cell. TU/ml value is usually 10- to 100-fold higher compared to pg/ml.

pg/ml gives you the amount of p24 capsid protein, which can be used to calculate the amount of viral particles. There are approximately 1 x 104 physical particles of lentivirus for every pg of p24 antigen.

What is a RCV test?

Cells that have been transduced with lentiviral particles should always be tested negative for replication competent viruses (RCV) before taking them to biosafety level 1 area. The test also ensures that your cell culture is virus-free, preventing biohazard environment (cross-contaminations and human users).

The test can be ordered through fugu-support(at)helsinki.fi. The service is provided by BVC unit and the service description can be found here.

How should I store viral particles ordered from FuGU Libraries?

Viral particles can be stored at -80C for up to one year. The infection activity is highest for fresh viruses and will decrease if the particles are frozen. However, most virus particles used for standard purposes can be frozen. Avoid thawing and freezing of the particles as this lowers and may destroy the infection activity.