4 results about "Multiple displacement amplification" patented technology

The application discloses a mutant PWT polymerase and a preparation method and application thereof, and relates to the technical field of biology. The mutant PWT polymerase has any one of A1) to A2): A1), an amino acid sequence shown in amino acids 407 to 986 of SEQ ID NO. 1; A2), an amino acid sequence obtained by connecting a tag to the N terminal or / and C terminal of the amino acid sequence shown in A1). The mutant PWT polymerase can obviously perform isothermal amplification under room temperature conditions and can tolerate a higher salt concentration, and is expected to become an ideal tool for rolling circle amplification and multiple displacement amplification.

A method of selective DNA amplification of a DNA mixture comprising a first population of DNA and a second population of DNA, wherein:—a) a catalytically dead Staphylococcus aureus Cas9 complex (dSaCas9) comprising a dSaCas9 protein or derivative thereof complexed with one or more guide RNAs having selective binding affinity for DNA sequences preferentially present in the first population of DNA compared to the second population of DNA is contacted with the DNA mixture under a first reaction condition, said first reaction condition being suitable for binding of the dSaCas9 complex to DNA sequences for which it has a binding affinity, and thenb) a strand-displacing DNA polymerase is contacted with the DNA mixture under a second reaction condition, said second reaction condition being suitable for amplification being suitable for amplification activity of the strand-displacing DNA polymerase; and related kits and uses.

Disclosed herein are mutants of bacteriophage Phi29 DNA polymerase with improved primer recognition, compared to the wild-type enzyme. Certain mutants comprise one or both of the mutations K64R or M97K. The provided mutants are capable of using more efficiently shorter and longer random synthetic DNA primers than wild-type Phi29 DNA polymerase, generating more amplification product in Multiple Displacement Amplification (MDA) reactions. The inventive mutants amplify human genomic DNA with less bias and better coverage in comparison to reactions carried out with wild-type Phi29 DNA polymerase.