NGS WORKFLOW

摘要

The present invention relates to improved semi-automated methods that permit the extraction of nucleic acids from samples, preparation of PCR and post-PCR preparation steps of DNA-libraries for next-generation sequencings methods that can be conducted. The methods and additional aspects relating to such methods are less laborious, safe costs, reagents and are less prone to contamination than comparable methods that are not automated.

主权项

1. A method of preparing a DNA library comprising the steps:
a) Extracting nucleic acids from a sample, b) Exposing the extracted nucleic acids to a mixture comprising UDG, a DNA polymerase and optionally a reverse transcriptase, and dUTP, c) Incubating the mixture to decontaminate the mixture from carry over amplification products derived from prior amplification reactions, d) Performing an amplification reaction in the presence of dUTP,wherein the decontamination reaction, optionally reverse transcription and DNA polymerase-conducted amplification reaction are performed in the same reaction mixture.