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Genome Biology Unit (GBU) Institute of Biotechnology, Viikki Tea Vallenius, Kaisa Laajanen, Niina Roine Genome scale reagents and services for functional.

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Presentation on theme: "Genome Biology Unit (GBU) Institute of Biotechnology, Viikki Tea Vallenius, Kaisa Laajanen, Niina Roine Genome scale reagents and services for functional."— Presentation transcript:

1 Genome Biology Unit (GBU) Institute of Biotechnology, Viikki Tea Vallenius, Kaisa Laajanen, Niina Roine Genome scale reagents and services for functional studies

2 Content General view Description of our libraries Basics of Gateway cloning How to order Clones and cloning service Future plans (e.g. kinome sets, custom projects) Genome scale reagents and services for functional studies

3 Genome scale reagents and services for functional studies

4 GBU services research groups of which 12 represent new groups in 2013 Genome scale reagents and services for functional studies

5 Value for scientists in Finland! - reduced costs - rapid delivery - easy access - screening possibilities Value for scientists in Finland! - reduced costs - rapid delivery - easy access - screening possibilities MGC Collection. cDNA library Human Mouse ORFeome Collaboration. Full ORFs Human Mouse cDNA/ORF sets available through Genome Biology Unit Genome scale reagents and services for functional studies

6 MGC collection = Mammalian Gene collection - National Institutes of Health initiative started The aim was to generate full-length protein coding cDNAs for human and mouse genes - sequence validated - MGC clones were intially obtained by random EST-screening of cDNA libraries ORF 5’ UTR3’ UTR - From the practical point of view: *clones we have are in various vectors -> not necessarily suitable for your studies immediately -> more time-consuming & expensive to transfer to other vectors *full length cDNA clones -> may contain 5’/3’ UTRs, which may have a regulatory role Genome scale reagents and services for functional studies

7 The ORFeome collection -ORFeome Collaboration, which aim is to provide sequence validated, ORF clones in Gateway entry vectors allowing easy transfer of the ORF sequences into various expression vectors -many available either with or without stop codons -From the practical point of view: *prior to expression studies need to be transferred into destination vectors *easy to transfer to any destination vector using Gateway cloning *contain only the protein coding sequences *GBU’s library has been intially validated ORF Genome scale reagents and services for functional studies

8 I. BP reaction = “ insert your gene of interest into the Gateway Technology” 2. negative selection 3. specific enzyme ORF 1.site-specific recombination kana Basics of the Gateway technology: Generation of an entry clone Genome scale reagents and services for functional studies

9 I. BP reaction = “ insert your gene of interest into the Gateway Technology” 2. negative selection 3. specific enzyme ORF 1.site-specific recombination kana Basics of the Gateway technology: Generation of an entry clone ORF entry clone Genome scale reagents and services for functional studies

10 2. LR reaction = “ transfer your gene of interest into destination vectors” 3. specific enzyme Basics of the Gateway technology: Generation of an expression clone 2. negative selection ORF 1.site-specific recombination amp 4. antibiotic selection Cloning service provided by GBU! = OFR library Genome scale reagents and services for functional studies

11 2. LR reaction = “ transfer your gene of interest into destination vectors” 3. specific enzyme Basics of the Gateway technology: Generation of an expression clone 2. negative selection ORF 1.site-specific recombination amp 4. antibiotic selection Several advantages: - rapid to perform - efficient : > 90% success rate - universal : no need to consider cloning sites - reversible : entry -> destination -> entry Several advantages: - rapid to perform - efficient : > 90% success rate - universal : no need to consider cloning sites - reversible : entry -> destination -> entry Genome scale reagents and services for functional studies

12 Content General view Description of our libraries Basics of Gateway cloning How to order Clones and cloning service Future plans (e.g. kinome sets, custom projects) Genome scale reagents and services for functional studies

13 Genome scale reagents and services for functional studies

14 Available clones on excel sheets Ordering form Sequence your clones easily – Collaboration with DNA sequencing and Genomics laboratory in Viikki and Biomedicum Sequencing Unit (BSU) Genome scale reagents and services for functional studies How to order: available ORF/cDNA sets

15 Available clones on excel sheets Genome scale reagents and services for functional studies

16 How to order: ordering form Available clones on excel sheets Ordering form Sequence your clones easily – Collaboration with DNA sequencing and Genomics laboratory in Viikki and Biomedicum Sequencing Unit (BSU) Genome scale reagents and services for functional studies

17 How to order: we recommend to verify you clones Available clones on excel sheets Ordering form Sequence your clones easily – Collaboration with DNA sequencing and Genomics laboratory in Viikki and Biomedicum Sequencing Unit (BSU) – Primers available for verification of ORF clones Genome scale reagents and services for functional studies Petri Auvinen Lars Paulin Tapio Tainola

18 ORF cloning service We clone your ORF clones of interest to multiple Gateway destination vectors Cloning format: Single clones or 96-well format Remember: FuGU can provide viral particle production service from your lenti-viral constructs Most commonly used Gateway destination vectors: N- terminal GST N- terminal 6xHis C- terminal V5 + 6xHis C- terminal GFP Lenti vector with C- term V5 Stop N-terminal emGFP + C- term V5, C-term emGFP N- and C- term. mCherry (a kind gift from Maria Vartiainen) Genome scale reagents and services for functional studies Mammalian Expression Bacterial Expression Insect Expression Entry Clone

19 96-well cloning and transfections trials Genome scale reagents and services for functional studies Pictures taken with Cell Insight in Light Microscopy Unit (LMU)

20 Genome scale reagents and services for functional studies GBU/FIMM/FuGU joined forces: -GBU is currently generating human Kinome ORFs (FIMM & FuGU have kinome knockdown reagents) Custom projects -Custom-tailored Gateway clonings -Primer script to design primers with Gateway tails -If you have ideas, we are happy to discuss Future Plans

21 Genome scale reagents and services for functional studies New service available - 3DHISTECH Pannoramic digital slide scanner

22 New service available Genome scale reagents and services for functional studies

23 Genome Biology Unit Contact information: Phone: Web pages: Personnel: Niina Roine M Sc Kaisa Laajanen M Sc Tea Vallenius MD, PhD Genome scale reagents and services for functional studies


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