Epigentek chromatin accessibility test kit principles and procedures

AI Meijie Epigentek EpiQuik The chromatin accessibility test kit is a complete set of optimization reagents , Designed to pass real-time  PCR  For nucleosomes of various biological samples / Gene specific analysis of chromatin accessibility by transcription factor mapping . It is a cost-effective and useful tool for rapid detection of open and closed chromatin .

AI Meijie Epigentek  EpiQuik Chromatin accessibility test kit features ：

1、J Its fast and convenient solution , Make the whole process （ From cell tissue samples to for  PCR  Ready to use  DNA） In a short span of  1  Hours  30  Done in minutes .

2、 It can be used for cultured cells as well as fresh and frozen tissues .

3、 The rapid process can minimize nuclear damage and the loss of separated chromatin components , Retain chromatin structure .

4、 The kit contains human internal control primers as a reference , Used to analyze samples  DNA  The degree of chromatin accessibility of specific gene targets in , And verify that proper enzymatic digestion has been achieved .

5、 Single reaction assay or high-throughput multi reaction assay can be selected , Make the measurement flexible .

Background information ：

The accessibility of regulatory elements in chromatin is crucial for many aspects of gene regulation . Nucleosomes located on regulatory elements inhibit transcription factors close to  DNA. To elucidate the interaction between chromatin and transcription factors , It is important to determine the accessibility of chromatin by mapping the location of nucleosomes along the genome . Generally speaking , The more condensed the chromatin , Transcription factors and others  DNA  The more inaccessible the binding protein is  DNA  And perform their tasks .DNA  The easier to approach , The more likely the surrounding genes are to be actively transcribed . The existence of nucleosomes （ Or not ） Directly or indirectly affect a variety of other cells and metabolic processes , For example, reorganization 、 Copy 、 Centromere formation and  DNA  Repair .

AI Meijie Epigentek EpiQuik Chromatin accessibility test kit principles and procedures ：

EpiQuik  The chromatin accessibility test kit contains real-time tests  PCR  For cells / Chromatin accessibility of tissue samples all necessary reagents for gene specific analysis . The kit includes internal control primers for determining the success of chromatin digestion . In this determination , Chromatin from cells / Separate from the tissue , And treated with nuclease mixture . Then use real-time  PCR  Isolation and amplification  DNA, To analyze the region specificity of chromatin accessibility . Chromatin status can be determined according to  DNA  To identify the accessibility of nuclease . External proteins （ Including exogenous nuclease ） Inaccessible to... In heterochromatin  DNA, Make it subject to nucleosome or  DNA/ Protection of protein complexes , And can be used in subsequent  PCR, Between digested and undigested samples  Ct  The change is insignificant . by comparison , In euchromatin  DNA（ Nucleosome depletion ） It can be accessed by exogenous nuclease , Make it susceptible to nuclease digestion , And because of the large difference between digested and undigested samples  Ct  Offset and cannot be used for  PCR.

Starting material and input quantity ：

Starting materials may include various mammalian tissue or cell samples , For example, cells from flask or microplate culture cells 、 Fresh and frozen tissue, etc . Each reactive cell / The amount of tissue can come from  1 x 10 5  individual Cells or 2 mg  Organization to  2 x 10 6  individual Cells or  40 mg  organization . For best response , The input dyeing quality should be about  0.5 x 10 6  individual Cells or  10 mg  organization .

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