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==Discussion== *'''Regardless which enzyme or condition was used, all dephosphorylated carriers are still ligated with adaptor in this test'''. **Even I did the inactivation step (70C 10min) following the End-repairing step, the situation was not improved. **B4-B6: De-P-Carrier prepared by SAP, 30min. **B5: De-P-Carrier prepared by SAP, 60min. **B6: De-P-Carrier prepared by ACP, 60min. *Possible reasons of the appearance of adaptor-ligated carrier: **The Dephosphorylation is not complete. **The De-P-Carriere was rephosphorylated after End-repairing. **The Phosphorylation "on templates (insert)" is not playing the critical roll for ligation. *The early size selection method is able to construct the library with only 10ng DNA (for both with or without carrier conditions). *The early size selection method can generate libraries '''with much less unspecific amplification'''.
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