Currently, we are really confused by a new function of Bst DNA ploymerase that has been commonly used for isothermal nucleic acid amplification. The phenomenon caused by the funtion seems like the self-multimerization of short double-stranded DNAs which, in theroy, should be independent and linear amplification products just like PCR’s products. The self-multimerization makes the products of isothermal amplification form genomic DNA-scaled fragments (over 10kb) with repeated sequences from the templates. Our established isothermal amplification only requires two linear primers (18-20 nt), one Bst DNA ploymerase, and the target region of template is below 100 bp. Through restriction enzyme digestion experiment, we are sure that the amplification is template-dependent, highly sensitive and specific. Additionally, previous studies have also reported the same phenomenon. However, due to the lackness of enzymology, we are frustrated by how to confirm the multimerization function of short double-stranded DNAs by Bst DNA ploymerse in molecular level, except massively sequencing the products. Look forwards to hearing from you. Thanks!
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