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The library was approximately 1e+9 clones with 24/24 unique clones having inserts at a ratio of almost 2 VH: 1VHH as judged by examining the amino acid composition of framework 2 [51].
Cloning efficiency was evaluated with PCR amplifications of bacterial colonies selected at random with vector specific primers and agarose gel analysis, resulting that 98% of colonies having inserts of variable size.
Of the remaining 378 clones with inserts, the inserts ranged in size from 9 to 292 kb, with 75% having inserts sizes 90 159 kb and 84% with inserts greater than 90 kb.
In addition, the arraying and screening of the library was facilitated by the high quality of the cDNA library itself which was reflected in all the clones (100%) having inserts, in the average insert size of 1.3 kb and in the transformation efficiency of 3.85 × 10.
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Recombinant phages having insert sizes of 200 bp were re-arrayed into new storage plates.
Only those colonies, having insert size between 1 kb to 3 kb were selected.
Four of the 122 clones (3.3%) had no insert and the remaining 118 (96.7%) all had inserts.
Out of 95 colonies, 71 (75%) had inserts.
Only 3% of the clones had inserts smaller than 500 bp (Fig. 1).
In addition, approximate 2.30% of the clones had inserts longer than 3.0 kb.
All cDNA clones used for developing EST resources have inserts consisting of complete 3'UTRs.
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