Quick conventional NGS DNA Library preparation for Illumina®
End repair and poly A addition in one step, simplified process and time saving
Total time : 3.0 hours
|Use fragmented DNA as the initial template :|
1. End repair and poly A addition in one step
Because DNA fragmentation does not result in homogeneous, blunt-ended fragments, end repair is needed to ensure that each molecule is free of overhangs, and contains 5' phosphate and 3' hydroxyl groups
Polyadenylation consists of the addition of a poly (A) tail, a succession of numerous adenosine (A) ribonucleotides at the 3 'end. This step facilitates the next step of linking the adapters.
2. Adapter ligation
Adaptors (short, double-stranded pieces of synthetic DNA) are then ligated to these fragments with the help of DNA ligase. The adaptors enable the sequence to become bound to a complementary counterpart. These adapters contain sequence motifs that are required for subsequent steps (clonal amplification and the actual sequencing).
3. Library amplification
Library amplification is required so that the received signal from the sequencer is strong enough to be detected accurately. This step is performed with a high fidelity DNA polymerase to avoid bias.
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