Thermo Scientific™ MuA Transposase

Description

• 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
• Thermo Scientific transposon products are based on the transposition machinery of the bacteriophage Mu
• During the lytic phase of the phage's life cycle the machinery replicates its genome by transposing repeatedly inside the host genome
• The Mu transposition reaction has been modified into an in vitro reaction catalyzed by a single enzyme - MuA Transposase
• In this system, one in vitro reaction is capable of generating more than a million transposon insertion clones
• Features The Mutation Generation System (MGS Kit) and Stop Generation System (STOP Kit) were developed for functional analysis of proteins
• These new transposon tools enable the creation of saturated libraries of mutated proteins in a single reaction with less hands-on time than any other method
• The location of the transposon insertion in each mutated clone can be mapped by either PCR or sequencing
• With MGS and STOP kits, thousands of mutated clones are ready for expression studies in just 2 to 3 days
• The MGS Kit contains the complete set of reagents for transposon-based linker scanning mutagenesis of any target protein
• The MGS Entranceposons are designed for making subtle changes in the structure of a target protein by inserting 15 bp in-frame linkers throughout the corresponding target gene
• This in-frame insertion allows for conservation of downstream sequences
• The STOP Kit Entranceposons 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 makes it possible to generate a saturated C-terminal deletion library from virtually any target protein with a maximum addition of three amino acids
• Highlights Efficient - Create saturated insertion libraries for sequencing and protein analysis in a single reaction Fast - Decrease hands-on time compared to conventional methods Random - Eliminate target site preference or insertion hot-spot Applications The STOP Kit generates truncated proteins for functional assays of: Enzymes Receptors Structural proteins etc
• 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 Includes MuA Transposase (F-750): MuA Transposase (0.22 μg/μL, 20 μL) 5x Reaction Buffer MuA Transposase concentrated (F-750C): MuA Transposase concentrated (1.1 μg/μL, 20 μL) 5x Reaction Buffer MuA Storage Buffer Stop Generation System (STOP Kit) (F-703): MuA Transposase (0.22 μg/μL in MuA storage buffer) Entranceposon (STOP-KanR) (100 ng/μL in TE buffer) 5x Reaction Buffer for MuA Transposase DMSO 100% Control Target DNA (370 ng/μL in TE buffer) MuEnd-2 Primer (25 μM in dd water) SeqE Primer (10 μM in dd water) SeqW Primer (10 μM in dd water) Mutation Generation System (MGS Kit) (F-701): MuA Transposase (0.22 μg/μL in MuA storage buffer) Entranceposon (M1-CamR) (100 ng/μL in TE buffer) Entranceposon (M1-KanR) (100 ng/μL in TE buffer) 5x Reaction Buffer for MuA Transposase Control Target DNA (370 ng/μL in TE buffer) 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