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We estimated the insertion time of all LTR-RTE families that harbor at least five intact copies.
The program automatically estimated the insertion size and direction between the paired reads and discarded the inferred incorrectly mapping pairs.
Using this rate, we have estimated the insertion time of 1,175 PbrCassandra, 188 MdCassandra, 194 PpCassandra, 69 PmCassandra, and 36 FvCassandra intact copies with TSDs.
Du et al. (2010) estimated the insertion dates of LTR retrotransposons using genetic distances calculated between left and right LTR sequences.
We estimated the insertion time for an element or the divergence time for elements of two different families following the strategy described by San Miguel et al. [ 45].
We estimated the insertion date of the complete copies using the method proposed by SanMiguel and co-workers [ 9], which is based on the divergence between the two LTRs of the copy.
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Since 1998 (SanMiguel et al. 1998), analysis of sequence divergence between paired LTRs from intact LTR-RTs, coupled with a molecular clock, has been the accepted method used to estimate the insertion times of LTR-RTs in plant genomes.
We proceeded to further validate our mutagenesis results and to estimate the insertion efficiency.
However, this method estimates the overall insertion date of the set of copies as a whole, and cannot estimate the insertion date for each individual genomic copy.
To overcome these caveats, we designed a new method (see Materials and Methods and Figure 1) to estimate the insertion date of the truncated copies.
MLEP (mec left extremity polymorphism) typing was performed to estimate the insertion of SCC Hg as per Ito et al. [ 15].
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