CD End Repair/dA-Tailing Module for Illumina, 5' phosphorylation, and dA-tailing are combined in a single step to transform fragmented DNA into 5-phosphorylated and 3-dAtailed DNA fragments, allowing direct ligation of Illumina sequencing adapters. It is an efficient enzyme mix for Illumina platform library construction that performs DNA end-repair and dA-tailing in one step. This kit can establish blunt ends on double-stranded DNA fragmented by covaries, chemical methods, or enzymatic methods.
Application:
Suitable for 100 pg-1 μg fragmented DNA. Used for one-step end repair, 5’ phosphorylated, and dA-tailing application to DNA library preparation for Illumina platform.
Storage:
The Mix should be stored at -30~-15°C.
Components:
Specifications:
| Sample amount | 100pg-1μg |
| Application | Suitable for 100pg-1μg fragmented DNA. Used for one-step end repair, 5’ phosphorylated, and dA-tailing application to DNA library preparation for Illumina platform. |
| Sample type | Fragmented dsDNA |
| Sequencing Platform | Illumina |
This step is for End Repair, 5' phosphorylated, and dA-tailing.
- Thaw the End Prep Mix 4 and spin down briefly. Prepare the reaction solution in a PCR tube as follows:
- Mix thoroughly by gently pipetting up and down. DO NOT Vortex! Spin down briefly.
- Put the tube in a PCR instrument and run the following PCR program (Hot Lid: 105°C):
- Adapter Ligation. Following the CD Universal Adapter Ligation Module for Illumina (MOD002) steps.
| Reagents | Volume of Sample |
| Input DNA | X µl |
| End Prep Mix 4 | 15 µl |
| ddH2O | Up to 65 µl |
| Temperature | Time |
| 20°C | 15 min |
| 65°C | 15 min |
| 4°C | Hold |
