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PCR was performed, with primer sequences generated against the genomic regions flanking the insert and a standard primer for the 3' end of the insertion sequence, to obtain homozygous insertion lines (Table S1).
A third approach, explored by Kim et al. [83], consists not in the direct milling of the end products of the conversion reaction (such as LiF/Fe in the 3/1 ratio, see Eq. 2) but in mimicking the composition at the end of the insertion reaction (such as LiF/FeF2 in the 1/1 ratio, see Eq. 1).
Apart from disrupting genes, IS6110 insertions have also been found to activate gene expression of nearby genes by an outward directed promoter (OP6110) at the 3' end of the insertion element [30], [31], [32].
Instead we have primer walked out from the defined 3' end of the insertion.
The amplitude of these peaks rose towards the end of the insertion.
Entries near the end of the insertion sheet were correlated with a larger number of mistakes.
Similar(32)
We noted that a 3-bp GCA motif was shared between the 5′ end of this insertion and the 5′ breakpoint in the TALEN spacer.
The variation was due to the presence of a 1471-nt inserrnA23SrnA23S (Figure 3), including 15-nt imperfect inverted repeats at both ends of the insertion and a 963-nt ORF.
Primers are normally designed for both 5' and 3' ends of the insertion element in order to target both sides of the insertion.
For accessions that carried the retrotransposon insertion as indicated by the Vrn-B1 marker, the 5′ and 3′ ends of the insertion were sequenced.
In areas 8 and 14 where the genomes differed as a result of an insertion event, direct repeats were identified at both ends of the insertion.
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