EpiNext ISAC Library Preparation Kit

The EpiNext™ ISAC Library Preparation Kit is a complete set of optimized reagents designed for, in a fast manner, the in situ accessible chromatin (ISAC) applications, including preparations of DNA library from low input samples to profile chromatin accessibility via next generation sequencing (NGS) using Illumina platforms. The kit has the following advantages and features:

• High sensitivity and flexibility: Can be used for both non-barcoded (singleplexed) and barcoded (multiplexed) DNA library preparation. The amount of input DNA can be as low as 1 ng with a range from 1 to 100 ng.
• Extremely fast and streamlined procedure: The procedure from input DNA to ready-to-use library is about only 1 hr. No clean-up is required between each step, and all reactions take place in the same tube before PCR, thereby saving time and preventing handling errors as well as loss of valuable samples.
• The most convenient for use: Contains all required components for each step of DNA library preparation, which are sufficient for end polishing, ligation, clean-up, size selection, and library amplification, thereby allowing the library preparation to be the most convenient with reliable and consistent results.
• Minimized bias: Ultra HiFi amplification enables researchers to reproducibly achieve high yields of DNA library with minimal sequence bias and low error rates.
  

Background Information
DNA library preparation is a critical step for NGS. For generating accurate sequencing data in NGS, the prepared library DNA should be sufficient in yield and high-quality. Also, as NGS technology continuously improves, DNA library preparation must be optimized accordingly. The amount of DNA prepared by the in situ accessible chromatin (ISAC) assay from low input cells often causes insufficient DNA library yields by using currently available library preparation methods. In addition, the currently used library preparation methods are time-consuming. To address this issue, EpigenTek developed the EpiNext™ ISAC Library Preparation Kit, specifically suitable for the prepared DNA from ISAC.

Principle & Procedure
This kit contains all reagents required at each step of the workflow for carrying out successful DNA library preparation with the use of prepared DNA from the ISAC assay. In the library preparation, the end repair/dA tailing (end polishing) of the prepared DNA is performed simultaneously. Adaptors are then ligated to both ends of the polished DNA fragments for amplification and sequencing. Ligated fragments are size-selected and purified with MQ beads, which allow quick and precise size selection of DNA. Size-selected DNA fragments are amplified with a high-fidelity PCR mix that ensures maximum yields from minimum amounts of starting material and provides highly accurate amplification of library DNA with low error rates and minimum bias.