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The mutants identified by these methods have a modified sequence of a sad gene that fails to pair with its homolog, thereby creating a self-silencing effect and compromising the silencing mechanism.
Alan Volkers, chairman of VH Bio Ltd said the company had no idea that the sequence they produced was a modified sequence of smallpox DNA.
The gfp- and lux-containing plasmids (pZEgfp and pSBluc) are derivatives of plasmids pZE1RM [ 17] and pSB377 [ 29], respectively, with a modified sequence of the multiple cloning site.
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Site-directed mutagenesis (SDM) methods are used to generate cloned DNAs with modified sequences for examining the importance of specific residues in protein structure and function.
Methylation independent PCR (MIP) primers for ABO were designed to amplify bisulfite modified sequences regardless of methylation status[34] and are ABO bisF (5'-ggatagggttttaaggtattagggTT-3') and ABO bisR (5'-gcaacacctcgAccatAActcc-3').
The signature of the different sequences – the reference sequence and the whole set of modified sequences – were calculated and compared by Euclidian metric in order to obtain distances to the reference.
Three SRL sequences were therefore synthesized: an unmodified SRL (used as a control for crystallization and diffraction) and two modified sequences with the introduction of a 2′-azido group into U2650 or A2670.
Expression of the modified sequences, either individually or in combination, in N. benthamiana leaves, resulted in the enzymes being directed to the chloroplasts.
The DSC3 promoter was amplified from the bisulfite-modified DNA by two rounds of PCR using nested primers specific to the bisulfite-modified sequence of the DSC3 CpG Island.
The copy numbers were calculated based on the linear regression obtained for a standard curve of 8 to 8 × 105 gene copy equivalents, using plasmid DNA containing the bisulphite-modified sequence of interest.
For both doping elements, a modified oxidation sequence of Ti was found, compared to the undoped TiAlN reference coating.
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