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Of the 38,419 PUTs (putative unique transcripts) longer than 150 bp in this reference assembly, 83.5% show similarity to ESTs from other spruce species and of the remaining PUTs, 3,704 show similarity to protein sequences from other plant species, leaving 4,167 PUTs with limited similarity to currently available plant proteins.
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However, this may be simply due to the difference in the study design, where up to 150 bp of the non-coding exon-intron boundaries were covered in this study compared to < 18 bp in the reference studies.
tRNA length ranged from 50 to 64 bp in the reference species "Metacrangonyx boveei".
These mtDNA sequences were each present at less than 100 bp in the reference nuclear genome (Table 1).
We chose microsatellites with a minimum of 20 repeat units of two to four bp in the reference sequence and designed amplification primers.
We sequenced the entire Ldmt gene (3,294 bp) in the reference strains and the clinical isolates for SNP analysis (5, 7 ).
In N50, both programs are outperformed by PacbioToCA [ 10], however, this is again due to a few very long repeats (approx. 5,000 bp) in the reference genome which were not resolved by GAML or Cerulean.
We note that our data set of indels does not include any insertions relative to the reference, because we only detect putative indels by having two read pairs aligned at positions separated by at least 1,000 bp in the reference genome (Materials and Methods).
Our close examination of the false-positive deletions showed that many of them correspond to the sites containing ambiguous 100 1000 N bps in the reference assembly as observed in our previous study.
The control sample was transfected with a gRNA that has no similar site (≥15 bp of 20 bp) in hg19 reference genome (Supplementary Fig. 1).
Exon regions shorter than 36 bp (in either reference) were excluded.
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