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Moreover, the cleavage products generated by crRNA and crHybs were identical, suggesting crHybs maintain the sequence specificity of Cas9 targeting (Supplementary Figs. 6 and 7).
Therefore, the temporary cluster head can change its state to a main cluster by selecting a subcarriers set that is not used by its adjacent clusters and trying its best to maintain the sequence of the subcarriers sets as c1, c2, c3.
This is known as purifying selection as potentially deleterious mutations are purged from the population in order to maintain the sequence of a functionally important site or region within the coding gene.
This demonstrates a negative purifying selection to maintain the sequence in both gene families.
To maintain the sequence depth uniformity of different fragments, a tight length range was selected (roughly 30-50 bp) and a pilot PCR amplification was performed to check the reduced representation library (RRL) features in this target length range.
The data presented here indicates that in these species there is a combination of strong pressure to maintain many cytochrome genes with weak pressure to maintain the sequence of most cytochrome genes.
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The kinetic of down-regulation of the GUS transcript was comparable to the one observed for the endogenous gene AtMYB60 [ 19], indicating that -619, -472 and -366 promoters maintain the sequences responsible for transcriptional down-regulation by ABA.
Their comfortable triumph also maintained the sequence of one-sided matches during the early stages of this year's competition with two of the other lowest scores in its history also being recorded - USA's 65 against Australia and Bangladesh's 93 against South Africa - and is bound to prompt further criticism of the format.
This is exemplified by the long term success of poliovirus vaccines, which has been related to purifying selection maintaining the sequence and hence structure of targeted surface proteins [38] conversely targeting the rapidly evolving proteins of HIV may have contributed to developing resistance [39].
DOI: http://dx.doi.org/10.7554/eLife.05290.007 We next performed site-directed mutagenesis to change the T found in high-plated fish to the G found in all sequenced low-plated fish, while maintaining the sequence of the high-plated marine haplotype throughout the rest of the enhancer construct.
We conservatively maintained the sequences that were confirmed in independent PCRs and those with at least three unique mutations for further analysis.
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