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With this range of inserts an appropriate library size for the screen was estimated after Clarke and Carbon.
And, in addition to that, the host expression machinery might be tuned to a broad range of inserts, with varying fragment types, origins and sizes.
Comparison of the size range of inserts observed in traditional and "gel-free" Fosill libraries indicates highly similar performance (mean insert size = 38.0 and 36.7 kb, respectively; Figure 2).
Due to a newly designed type of polylinker, directional cloning of a wide range of inserts is greatly facilitated, and HA-tagging of the insert of interest is possible if needed, depending upon which pPRIG vector is selected.
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Prior to the arrival of NGS platforms, WGS on larger genomes was possible because Sanger-based sequencing platforms generated paired-end reads of 700 bp or greater and a range of insert sizes from 3 to 40 kb (Weber and Myers 1997).
Its major feature was broad range of insert size and very low fraction of PE reads.
In consequence, it remains unclear, if the sub-libraries actually contained the expected range of insert sizes.
A Colony-PCR was performed to assess the range of insert sizes as well as the cloning efficiency.
A big insert of 465 nt in the middle of the cluster created no putative tRNA structure in the range of insert.
Using the robust Taq enzyme or a proofreading Pfu DNA polymerase mutant, the method is applicable to a broad range of insert sequences.
This may be attributed to the size range of insert, the restriction enzyme used for genomic DNA library construction and the approach used for SSR enrichment, etc. [ 7].
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