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In silico conversion involved converting all cytosines to thymines.
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In order to identify CpG and non-CpG methylation sites, B-SOLANA aligns bisulfite sequences to two in- silico conversions of the reference genome (Supplementary Fig. 2).
Due to the two-base encoding of SOLiD sequencing, in silico conversions of any residual bisulfite read cytosines into thymines, which can be carried out in basespace data to avoid bisulfite-mismatches during alignment, cannot be performed on bisulfite colorspace sequences, because sequencing errors would lead to the incorrect translation of colorspace to basespace (Supplementary Fig. 1).
We found a significant difference between in vitro SNP and in silico SNP conversion rates, a lower rate being observed for in silico SNPs.
Satellite 1 sequences were extracted and subjected to in silico bisulfite conversion based on their genomic coordinates in the UCSC database.
There he learned that "in silico" they were test-driving vehicles, running them through scenarios in simulation before testing them on the track or the open road.
BS aligners employing three-letter approaches perform in silico C-to-T conversion for both reads and reference sequences prior to mapping.
The candidate group of HCGs had the highest successful rate for conversion of in-silico SNP loci into SNP markers (61.1%) while this rate was only 40.1% for the candidate group of RGAs.
It is possible that the rate of conversion of in-silico SNPs can be improved even more in WWP by optimizing primer design and PCR amplification conditions because we found that some iPlex failed SNPs could be genotyped properly by qPCR genotyping methods such TaqMan and HRM (Liu, unpublished data).
SNP marker conversion rates from in-silico SNPs to validated loci have been reported for maritime pine (P. pinaster) (42.5%), lodgepole pine (P. contorta var. latifolia) (30.0%), Aleppo pine (P. halepensis Mill).
Variation in SNP marker conversion rates suggests criteria for in-silico SNP selection and genotyping design, as well as types of genotyping platforms are important.
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