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A mutant sequence containing 9 mutations was also manufactured as a control to ensure specificity of knockdown.
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T = True mutants that sequences contain mutations confirmed by Mutation Surveyor®.
The mutants that are modeled the worst are surprisingly not the heavy mutant sequence 7 (which contains a number of 13 total mutations), but rather sequence 2, and 6.
The span of a motif refers to the number of sequences containing its d-mutant.
Plasmids containing mutant sequence served as the template for the analytical sensitivity test.
Our results showed that reporter plasmid containing the miR-183 targeting sequence of ITGB1 mRNA displayed a significantly decreased luciferase activity in cells transfected with miR-183 (P < 0.05), whereas luciferase activity of reporter plasmid containing mutant sequence of ITGB1 was not changed following forced expression of miR-183).
One of the oligonucleotide pools (LE) contained the lox71 left arm mutant sequence and the other (RE) contained the lox66 right arm mutant sequence [ 12].
One of the two oligonucleotide pools (LE) contained the lox71 left arm mutant sequence and the other (RE) contained the lox66 right arm mutant sequence [ 12].
Genes containing the mutant sequence with a C→A substitution at the serine 27 codon do not contain the restriction enzyme site and, therefore are not cut by the enzyme.
Hybridisation to the array is then performed using a two-colour system to provide a direct comparison between wild-type sequence and the test sample potentially containing a mutant sequence.
Meanwhile, downregulation of miR-100 could lead to a significant increase in luciferase activity the reporter contained a wild-type sequence (WT), but not when it contained a mutant sequence (MT) within the miR-100 binding site.
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