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Creative BioMart's strong expertise in cloning and subcloning allows us to accelerate your research project by providing gene cloning and subcloning. We will design personalized primers and use the most appropriate PCR method for fragment amplification based on the actual condition of your target gene in order to help you reveal or probe the function of a specific gene.
Gene cloning refers to the use of specific primers to obtain the DNA sequence of the gene of interest from the sample tissue or cell by PCR technology, and then clone it into a specific vector and transfer it into the cell to obtain a large number of copies. Obtaining gene sequences and specific DNA sequences is an important basis for studying DNA structure and function in molecular biology research.
Fig 1. The flowchart of SPI cloning. (Reddy, T. R, et al., 2015)
Gene subcloning is to cut the gene of interest or a specific DNA fragment with enzymes or re-PCR amplification and clone it from one vector into another vector. This method enables the observation of the gene expression process and exact function. The cloning and subcloning of genes provide an experimental basis for genetic engineering research or molecular biology research.
Fig 2. The flowchart of subcloning DNA fragments from genomic DNA. (Sharan, S. K, et al., 2009)
Creative BioMart provides professional and rapid gene cloning and subcloning services including but not limited to the following to meet your research needs.
We will design scientifically reasonable primer fragments based on the gene sequence you provide and obtain the gene fragments you need by PCR. Or find out the DNA sequence of the gene encoding the protein according to the protein sequence (for example, clone the gene using a specific antibody). Various other methods including TOPO cloning, Gateway cloning, TA cloning, GeneArt seamless cloning, blunt-end cloning, long fragment cloning, etc. can also help us obtain your target gene fragments. We can assemble multiple DNA fragments simultaneously for you, and avoid multiple rounds of restriction enzyme digestion, DNA end-repair, dephosphorylation, ligase ligation, enzyme inactivation, and DNA sample loss, which can save a lot of experimental time.
The probes can be used as radioactive or non-radioactive markers, then hybridized with DNA treated with different endonucleases, and finally, the identified fragments are cut off from the gel and cloned into specific vectors (plasmids, phages, or viruses) for sequence determination or gene function analysis.
Gene cloning of unknown sequences by random primer method, differential display PCR (DD- PCR), and representative difference analysis PCR (RDA- PCR).
The customer provides the PCR product of the amplified target gene and we insert the PCR product into the specific expression vector.
We will design a plan according to the nature and structure of the vector you need to modify, and we can complete the modification of MCS of existing vector polyclonal sites, and the modification of functional elements such as promoters, enhancers, and screening markers of existing vectors. Then we will subclone the gene fragments or functional elements into each vector according to your requirements.
If you have any special requirements for our cloning and subcloning service, please feel free to contact us. We look forward to working with you on attractive cloning and subcloning projects.
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