CUT&Tag PCR Master Mix #63228
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Product Information
Storage
Store at -20ºC. This product is stable for 18 months if stored properly.
Protocol
Product Description
The CUT&Tag PCR Master Mix is an optimized 2X reaction mix for PCR amplification and DNA library preparation for CUT&Tag DNA samples. The non hot-start DNA polymerase in this product ensures the success of gap filling extension for tagmentated DNA. This master mix formulation is supplied at 2X concentration and contains all PCR components required for amplification of DNA, except primers and a DNA template. This product is provided in 840 μL volumes sufficient for preparation of 24 PCR reactions, and is compatible with CUT&Tag DNA sample generated by CUT&Tag Assay Kit #77552 or CUT&Tag pAG-Tn5 (Loaded) #79561 and the index primers provided in the CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. This product is not compatible with library preparation for DNA samples from SimpleChIP® Chromatin IP Kits (#9003, #9005, #56383) or the CUT&RUN Assay Kit #86652.
Specificity / Sensitivity
Species Reactivity:
All Species Expected
Background
Similar to Cleavage Under Targets and Release Using Nuclease (CUT&RUN), Cleavage Under Targets and Tagmentation (CUT&Tag) is a powerful technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1-3). CUT&Tag has many of the same advantages as the CUT&RUN assay in that it provides a rapid, robust, and true low cell number protocol for detection of protein-DNA interactions in the cell. In addition, the CUT&Tag assay adds an in situ adaptor DNA ligation step carried out by the pAG-Tn5 enzyme, in which an adaptor DNA is ligated directly to antibody-targeted chromatin DNA fragments in the cell. As a result, subsequent DNA library preparation is much faster and easier than library preparation following the CUT&RUN assay, free from DNA end repair, A-tailing, and adaptor ligation in vitro. CUT&Tag works very well for analyzing histone modifications, in addition to mapping some transcription factor and cofactor binding.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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