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                          TagMaster® Site-Directed Mutagenesis Kit
                                              — Easily insert/switch any tag into your gene

GM7001  TagMaster® Site-Directed Mutagenesis Kit
  !!!This item has to be shipped overnight!!! 		20 rxn $660.00

GM7002  TagMaster® Site-Directed Mutagenesis Kit
  !!!This item has to be shipped overnight!!!		 6 rxn $399.00


The kit provides a rapid, efficient and accurate method for large fragment mutagenesis in a single reaction. It has been optimized for mutagensis of plasmids of up to 10kb. In addition to point mutation, the kit allows the highest accuracy and efficiency to introduce a variety of mutation: insertion/substitution(~60bp), deletion(~660bp) and multiple sites mutation (two sites). 
It is the first of the kind in the world that can insert/switch tags such as 6HIs, Myc, Flag, HA V5  into a gene by a simple single reaction.  CRISPR/Cas9 expression plasmid can be easily constructed by switch/insertion guide RNA sequence in a single reation.
      
      Highlights:
  • Powerful  -insertion/substitution (1~60bp)
                  -insert/switch tags( 6His, Myc, Flag, HA, V5......)
                  -insert/switch sgRNA sequence in CRISPR/Cas9 plasmid
                  -deletion (1~660bp)
                  -multiple sites mutations (2 sites)
                  -suitable for plasmid up to 10kb
  • Efficient: 80%~99% mutation rate
  • Accurate  -high-fidelity enzyme blend
                   -linear DNA amplification, non-PCR procedure
  • Fast and simple  -easy 2-step protocol
                             -1h~2h reaction time

 

      Overview of TagMaster Mutagenesis Method:
 
The TagMaster Mutagenesis Kit has unparalleled efficiency. As shown in Fig 1, 99.4% and 98.7% mutation rates have been achieved for a 2bp substitution and 18bp(6His tag) insertion.

The TagMaster mutagenesis technology circumvents the limitations of traditional restriction enzyme based cloning, enabling you to easily obtain different tags for a gene. As shown in Fig 2, different tags (6His, HA, Myc+Flag, Myc, Flag) had been successfully inserted into a 7.4kb plasmid. 

The kit has successfully deleted GST Gene (660bp) from a GST expression plasmid. Deletion of longer fragment may work as well, although it has not been tested yet.

The proprietary TagMaster competent cells in the kit achieved significant higher mutagenesis rate than other cells, such as DH5α(Fig 3).

 

  

Fig 1. Unparalleled mutagenesis rate achieved by TagMaster Site-Directed Mutagenesis Kit. 
Three mutagenesis reactions were performed under conditions of 95°C for 5min, followed by 22 cycles of (95°C 20sec, 60°C 10sec, 68°C 2min). Without any treatment, the products of reactions were directly transformed into TagMaster competent cells and spread onto LB plates. Clone number in each plate were counted as shown in panel A. The mutagenesis rates were calculated and results are shown in panel B. Please click the figure for details.

 

Fig 2. Successful insertion of different tags (6His, HA, Myc+Flag, Myc, Flag) into a 7.4kb plasmid.
The Mutagenesis reactions were performed under conditions of 95°C 5min, 20cycles of (95°C 20sec, 60°C10sec, 70°C 3min20sec). The reaction products were run in 1% agrose gel (as shown above). The products were then directly transformed into TagMaster competent cells. The insertion of tags was verified by sequencing. The successful rates of all 6 mutagenesis reactions were 82%~91%. Please click the figure for details.

 

Fig 3. TagMaster competent cells achieve significant higher mutagenesis efficiency than DH5α competent cells. Please click the figure for details.

 Ordering Information
Cat# Description Price Document Order
GM7001  TagMaster® Site-Directed Mutagenesis Kit, (20 rxn)
 (For Academic Use Only)		 $660 Protocol
MSDS		 Place order
GM7002  TagMaster® Site-Directed Mutagenesis Kit, (6 rxn) Tial
 (For Academic Use Only)		 $399 Protocol
MSDS		 Place order
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