The Universal DNA Library Prep Kit for Illumina offers a quick and easy workflow for library preparation in Illumina-based next-generation sequencing (NGS) applications. This kit allows library preparation from as little as 100pg starting material and enables the construction of a single library in just 75 minutes. The Kit can not only modify the operation process and shorten experiment time, but it can also reduce the risk of contamination as an improved version with fewer handling steps and cleanup steps. The Kit allows for a higher library conversion rate and more consistent coverage yields by enhancing adapter ligation efficiency and reducing PCR amplification bias. This kit can be used to make PCR or PCR-free libraries from a variety of input DNA and is also compatible with target capture systems. All Kit components are subjected to rigorous quality control to ensure that library preparation is consistent and repeatable. This kit is also compatible with a variety of other products.
Storage:
All components should be stored at -20℃
Components:
Specifications:
| Sample amount | 100 pg - 4 μg |
| Features | Fast and convenient workflow enables a single library preparation within 75min. Fewer handling steps and fewer cleanup steps. Enables higher library conversion rate, higher and uniform coverage yields. |
| Application | It is recommended to use this kit for: Whole genome sequencing Whole exome or targeted sequencing (using Roche NimbleGen TM SeqCapTM EZ, Agilent SureSelect, Illumina TruSeq, or IDT xGen TM Lockdown TM Probes or other hybridization capture systems) Amplicon sequencing ChIP-seq Metagenome sequencing Methylation sequencing |
| Species Category | Compatible with any species |
| Sample type | genomic DNA, cell-freeDNA (cfDNA), circulating tumor DNA (ctDNA), formalin-fixed paraffin-embedded DNA (FFPE DNA), Chromatin immuno-precipitation DNA (ChIP DNA) and Amplicons |
| Sequencing Platform | Illumina |
Workflow
Protocol
Step 1. End Preparation
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.
- 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.
Step 2. Adapter Ligation
This process is to ligate End Preparation products to Adapters.
- Dilute adapter stocks to the appropriate concentration.
- Thaw the Rapid Ligation Buffer and mix thoroughly. Place on ice.
- Prepare the reaction solution in a sterile PCR tube.
- 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.
- Cleanup of Adapter Ligation products with CD DNA Clean Beads.
Step 3. Library Amplification
This process is to amplify the purified or size-selected adapter ligation products. Whether to proceed with this step depends on the amount of input DNA, whether adapters are in complete length, and the need of application. If adapters are not in complete length (i.e. DDI001/DDI002), this step is necessary. If adapters are in complete length (i.e. DSI001/ DSI002), for input DNA < 50 ng, amplification is recommended. Skip this step if input DNA is ≥ 50 ng or there is no need for library amplification.
- Thaw PCR Primer Mix 3 and HiFi Amplification Mix, and mix thoroughly. Prepare the reaction solution in a sterile PCR tube.
- 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.
- For size selection, please refer to Appendix 1. If there is no need for size selection, please purify the products with CD DNA Clean Beads.
Step 4. Quality Control of Library
