可以通过 qPCR 分析 CUT&Tag 富集的 DNA，还是必须进行 NGS 来分析我的数据？

If qPCR analysis is desired, we recommend performing CUT&RUN, as CUT&Tag DNA is not compatible with qPCR. 在 CUT&Tag 实验步骤中 标签化 DNA 增溶解所要求的 58℃ 时样品孵育步骤打开核膜，从而导致最终的 CUT&Tag DNA 样品由靶标结合的标签化 DNA 和背景基因组 DNA 组成。如果用针对最终 CUT&Tag DNA 样品上靶标基因的引物进行 qPCR，则无法区分 CUT&Tag 信号与背景信号。

However, qPCR analysis of target genes is possible using a CUT&Tag DNA library. This is because, during the library amplification process, the tagmented DNA fragments are selectively enriched while the background genomic DNA is diluted out. The qPCR analysis on a CUT&Tag DNA library can serve as a QC step before next-generation sequencing (NGS). We recommend using the CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415 for CUT&Tag DNA library preparation.

More information on this topic can be found on the CUT&Tag Frequently Asked Questions page. 

Last updated: 2024 年 9 月 12 日