Mutation Generation System Kit

Finnzymes was recently acquired by Thermo Fisher Scientific, and this line of products is now part of the company's Thermo Scientific PCR portfolio. For customers in North America, you can also place your Finnzymes products orders through www.dharmacon.com and benefit from faster delivery times Description The Mutation Generation System Kit (MGS Kit) is designed for rapid construction of insertion mutation libraries for any kind of DNA clones. The system employs the highly efficient transposition machinery of the bacteriophage Mu to generate a pool of 15 bp insertion mutants that can be utilized in a variety of functional analyses of the encoded proteins or regulatory DNA regions. Applications The MGS Kit generates random fifteen basepair in vitro insertions into any target DNA for: Rapid generation of in-frame five amino acid insertion libraries of any protein for functional analyses Rapid and random mutagenesis of cloned promoters and other regulatory DNA regions Random insertion of a NotI restriction enzyme site into any target DNA clone Advantages Thousands of different insertion clones from a single reaction Generates random insertions of 5 amino acids in all 3 reading frames Short in-frame insertions; no stop codons Flexibility in mapping mutants of interest: mutations are easily mapped by NotI or PCR*. Faster and more effective than linker scanning mutagenesis Components The kit contains sufficient materials for 10 reactions: MuA Transposase: 10 µl (0.22 µg/µl in MuA storage buffer) Entranceposon (M1-CamR): 10 µl (100 ng/µl in TE buffer) Entranceposon (M1-KanR): 10 µl (100 ng/µl in TE buffer) 5x Reaction Buffer for MuA Transposase: 100 µl Control Target DNA: 10 µl (370 ng/µl in TE buffer) NotI Miniprimer: 50 µl (25 µM in dd water) Storage Store the components at –20 °C. more details »

Template Generation System II Kit

Finnzymes was recently acquired by Thermo Fisher Scientific, and this line of products is now part of the company's Thermo Scientific PCR portfolio. For customers in North America, you can also place your Finnzymes products orders through www.dharmacon.com and benefit from faster delivery times Description Transposon technology is the first technology that allows scientist to insert constructed linear DNA fragment into circular or linear DNA target randomly and very efficiently. The Template Generation System™ II Kit (TGS™ II Kit) is based on the transposition reaction of the bacteriophage Mu. This system has been simplified to work in vitro, and an artificial Mu transposon, designated as the Entranceposon, has been constructed. The reaction is catalyzed by a single enzyme, MuA Transposase. TGS II Kit provides the tools for inserting the Entranceposon into foreign DNA at random locations. Insertion of an Entranceposon into unknown target DNA provides primer binding sites for different applications. A simple and fast in vitro transposition reaction takes place in a single reaction tube. All components needed as well as detailed instruction manual to perform the reactions, are included in the kit. TGS II Kit is an updated version of the TGS™ I Kit. Applications The Template Generation System™ II Kit is designed for inserting an artificial Mu transposon into unknown target DNA to provide primer binding sites for: Rapid sequencing of templates without primer walking or custom primers Shotgun sequencing of large DNA clones directly without time-consuming fragmentation and subcloning into a library Insertion of PCR priming sites into target DNA with an unknown sequence Random insertional mutagenesis of cloned DNA; direct amplification, mapping or sequencing using primers at the insertion site Special Applications: BAC sequencing Transposition reaction into linear target DNA Advantages Provides templates for DNA sequencing more simply and with less hands-on time than any other method Thousands of ready-to-sequence templates from a single transposition reaction: Enough to sequence even large DNA clones (e.g. BAC clones) and without fragmentation and subcloning Simple mapping of the Entranceposon insertions by colony-PCR or by restriction enzyme digestion enables directed sequencing. Consequently, fewer sequencing reactions are required to complete a given sequence than with the "shotgun" approach Universal primers: The sequencing and mapping primers are included in the kit, no need for custom primers. Bidirectional sequencing: A single template clone can be used for sequencing the flanking DNA on both sides of the Entranceposon insertion Tree different antibiotic resistance genes available in each TGS II Kit Kit Components See our list of Kit Components Storage Store the components at -20 °C. more details »

Transposon-based STOP™ Kit

Finnzymes was recently acquired by Thermo Fisher Scientific, and this line of products is now part of the company's Thermo Scientific PCR portfolio. For customers in North America, you can also place your Finnzymes products orders through www.dharmacon.com and benefit from faster delivery times Description Transposon-based STOP™ Kit is developed for functional analysis of proteins. This transposon tool enables the creation of saturated libraries of mutated proteins in a single reaction. The Entranceposons in the STOP Kit contain translational stop codons in all three reading frames within the terminal portion of the transposon sequence. The proprietary modification of the Stop Generation System™ Kit makes it possible to generate a saturated C-terminal deletion library from virtually any target protein with a maximum addition of three amino acids. The transposon-based method requires less hands-on time than any other method, thousands of mutated clones are ready for expression studies in just two days. Applications The STOP Kit generates truncated proteins for functional assays of: Enzymes Receptors Structural proteins etc. Transposition reaction into linear target DNA Advantages Saturated library of truncated proteins from a single reaction in two days Translational STOP codon in all three reading frames The target DNA sequence can be unknown Faster and more effective than conventional methods No specific primers required Components The kit contains sufficient materials for 10 reactions. MuA Transposase: 10 µl (0.22 µg/µl in MuA storage buffer) Entranceposon (STOP-KanR): 10 µl (100 ng/µl in TE buffer) 5x Reaction Buffer for MuA Transposase: 100 µl DMSO 100 %: 500 µl Control Target DNA: 10 µl (370 ng/µl in TE buffer) MuEnd-2 Primer: 50 µl (25 µM in dd water) SeqE Primer: 250 ul (10 uM in dd water) SeqW Primer: 250 ul (10 uM in dd water) Storage Store the components at –20 °C. more details »

Transposon products available separately

Finnzymes was recently acquired by Thermo Fisher Scientific, and this line of products is now part of the company's Thermo Scientific PCR portfolio. For customers in North America, you can also place your Finnzymes products orders through www.dharmacon.com and benefit from faster delivery times Description MuA Transposase F-750: MuA Transposase 0.22 µg/µl (20 µl), 5x Reaction Buffer (100 µl) F-750C: MuA Transposase concentrated 1.1 µg/µl (20 µl), 5x Reaction Buffer (100 µl), MuA Storage Buffer (100 µl) Entranceposons with wild type MuA recognition sites, with a polylinker: F-764 Entranceposon (TetR): The marker gene tet confers resistance to tetracycline. F-771 Entranceposon (supF): The marker gene supF codes for mutated tRNA that recognizes the amber stop codon (UAG) and can suppress a nonsense mutation in an E. coli strain. If the strain carries an amber nonsense mutation in a gene that is required for growth, for example an antibiotic resistance marker in a plasmid,the suppressing tRNA can insert tyrosine at the UAG stop codon, resulting in a full length gene product. Entranceposons with wild type MuA recognition sites,without the polylinker: F-778 Entranceposon (CamR-3): The marker gene cat confers resistance to chloramphenicol. F-779 Entranceposon (KanR-3): The marker gene npt confers resistance to kanamycin. F-784 Entranceposon (TetR-3): The marker gene tet confers resistance to tetracycline. F-772 Entranceposon (supF-2) (See above) F-774 Entranceposon (lacZ): The marker gene lacZα codes for the α-polypeptide of ß-galactosidase. Expression of the marker can be utilized for blue-white selection of the insertion clones. F-775 Entranceposon (lacZ PT): Promoter Trap Entranceposon for searching transcribed regions in target DNA. Expression of the promoterless marker gene lacZa requires transcription from a promoter in the target DNA upstream from the insertion site. Plasmid Entranceposons F-765 pEntranceposon (CamR): Entranceposon (CamR)(F-751) in pUC19 F-766 pEntranceposon (KanR): Entranceposon (KanR)(F-759) in pUC19 F-767 pEntranceposon (TetR): Entranceposon (TetR)(F-764) in pUC19 F-773 pEntranceposon (supF): Entranceposon (supF)(F-771) in pUC19 more details »