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For family classification, the biological property of interest (sequences that share common ancestry) corresponds to a precise mathematical construct (cliques).
The resulting cDNA was amplified in SybrGreen real-time quantitative PCR assays using validated primers specific for the genes of interest (sequences are given in Supplementary Table 2).
Q-RT-PCR was performed on the cDNA using iQ SYBR Green Supermix (Bio-Rad) and respective primers for the genes of interest (sequences available upon request) as per the manufacturer's recommendations.
Cultivation of these transfected cells in methotrexate (MTX) containing medium results in amplification of the dhfr gene and gene of interest sequences and thus high-level expression of the gene of interest.
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The sensitivity would depend on the length of the region of interest, sequencing depth, and fetal DNA fraction.
Clone λz3-79 conthening the longest insert was subcloned and subclones of interest sequenced confirming that it did not contain sequences from region B (Figure 1).
It is then necessary to use other techniques through epigenetic analyses (markers of promoter regions or entire gene), targeted transcripts of interest sequencing after capture on a DNA chip (capture-RNAseq) and 5′ and 3′ extremities identification analyses [ 8– 11].
All constructs confirmed to be the sequence of interest by sequencing.
Direct purified PCR products or gel-purified bands of interest were sequenced using Sanger sequencing method by Beckman Coulter Genomics facility (Takeley, United Kingdom).
In most cases of interest, these sequences converge to their limits extremely slowly, or even diverge.
In order to identify genes or functions in an organism of interest, genome sequences can be produced and analyzed using comparative genomics tools.
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