ABSTRACT. Site-directed mutagenesis is an essential technique for investigating the mechanisms of gene regulation on a molecular level, as well as for exploring post-translational modifications and functional structure at the protein level. Polymerase chain reaction combining in vitro synthesis of oligonucleotide primers allows for site-directed mutation to be performed with ease. However, site-directed mutagenesis is difficult when larger plasmids are involved. Here, we present a novel method for generating large gene site-directed mutagenesis products based on a cut-and-paste-based cloning strategy. This method uses 4 primers, incorporating relevant mutations and restriction enzyme site sequences, to generate 2 DNA fragments by polymerase chain reaction. The fragments are then ligated into TA cloning vectors. Large genes containing mutations of interest were obtained by cutting and then pasting, and then inserting one fragment into another T-vector. We demonstrated the practicality of this method by creating a G59S mutation within the p150 Glued -encoding gene.

crystals down to few layers and even monolayer. MoTe 2 is a typical member among TMDCs, hosting fascinating electronic properties, such as extremely large magnetoresistance, [6,7] the emergence of superconductivity, [8] the existence of topological Weyl nodes [9][10][11][12] and semiconducting behavior with a finite band gap. [13] Layered structure with weak interlayer coupling allows for the change of layer stacking order and subsequently gives rise to a structural polymorphism. [14][15][16] In fact, MoTe 2 appears in three kinds of structures: the trigonal prismatic coordinated 2H phase (space group P6 3 /mmc), distorted octahedral coordinated structures, including the monoclinic 1T′ (space group P2 1 /m), and the orthorhombic T d (space group Pmn2 1 ) phases. The 2H phase exhibits semiconducting behavior, while 1T′ and T d phases show semimetallic characters. [13,17,18] As shown in Figure 1a, at room temperature, distortion of in-plane bond give rise to monoclinic unit cell 1T′ phase (tilt angle of ≈93.9°). When decreasing the temperature below 250 K, the shift of layer stacking results an orthorhombic structure. [16,19] In its monoclinic 1T′ phase, monolayer MoTe 2 has been proposed to be topological insulator exhibiting quantum spin Hall effect. [3] While the T d phase demonstrates a number of unique properties, such as extremely large magnetoresistance, nontrivial superconductivity, [20][21][22] switchable ferroelectricity, giant nonlinear Hall effect, and unconventional planar spin Weyl semimetal T d -MoTe 2 has recently attracted much attention due to its intriguing electronic properties and potential applications in spintronics. Here, Fe-intercalated T d -Fe x MoTe 2 single crystals (0 < x < 0.15 ) are grown successfully. The electrical and thermoelectric transport results consistently demonstrate that the phase transition temperature T S is gradually suppressed with increasing x. Theoretical calculation suggests that the increased energy of the T d phase, enhanced transition barrier, and more occupied bands in 1T′ phase is responsible for the suppression in T S . In addition, a ρ α -lnT behavior induced by Kondo effect is observed with x ≥ 0.08, due to the coupling between conduction carriers and the local magnetic moments of intercalated Fe atoms. For T d -Fe 0.15 MoTe 2 , a spin-glass transition occurs at ≈10 K. The calculated band structure of T d -Fe 0.25 MoTe 2 shows that two flat bands exist near the Fermi level, which are mainly contributed by the d yz and d x y 2 2 − orbitals of the Fe atoms. Finally, the electronic phase diagram of T d -Fe x MoTe 2 is established for the first time. This work provides a new route to control the structural instability and explore exotic electronic states for transition-metal dichalcogenides.