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• W2156865345 abstract "Rolling circle amplification (RCA) of plasmid or genomic DNA using random hexamers and bacteriophage phi29 DNA polymerase has become increasingly popular in the amplification of template DNA in DNA sequencing. We have found that the mutant protein of single-stranded DNA binding protein (SSB) from Thermus thermophilus ( Tth ) HB8 enhances the efficiency of amplification of DNA templates. In addition, the Tth SSB mutant protein increased the specificity of phi29 DNA polymerase. We have overexpressed the native and mutant forms of Tth SSB protein in Escherichia coli and purified them to homogeneity. In vitro , these proteins were found to bind specifically to single-stranded DNA. Addition of Tth SSB mutant protein to RCA halved the elongation time required for phi29 DNA polymerase to synthesize DNA fragments in RCA. Furthermore, the presence of the Tth SSB mutant protein essentially eliminates nonspecific DNA products in RCA reactions." @default.
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• W2156865345 date "2006-05-17" @default.
• W2156865345 modified "2023-09-25" @default.
• W2156865345 title "Improvements of rolling circle amplification (RCA) efficiency and accuracy using Thermus thermophilus SSB mutant protein" @default.
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• W2156865345 doi "https://doi.org/10.1093/nar/gkl350" @default.
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