Conventional NGS DNA Library Preparation Kits for Illumina®
Classical process with efficiency optimization
Total time : 3.5 hours with PCR and 3.0 hours without PCR
|Use fragmented DNA as the initial template :|
1. Repair ends and phosphorylate the 5' end
DNA is fragmented either enzymatically or by sonication to create smaller strands
2. Add dA-tail to the repaired products
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.
3. 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).
4. Library amplification
Library amplification is required so that the received signal from the sequencer is strong enough to be detected accurately. The Illumina technology uses the "Bridge PCR" method.
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