NovoNGS® ChiTag® pAG-Transposase(Cat. No.:M058-YH01)

Home > Product > NovoNGS® ChiTag® pAG-Transposase

COA

Price：

$234

SKU：

M058-YH01-01A

Size：

12 Rxns

Count：

Information

Related Products

Citations

Troubleshooting

Description

NovoNGS® ChiTag® enzyme has the activity of breaking double-stranded DNA and binding to antibodies. Based on its function, we applied it to CUT&Tag for protein-DNA interaction research. CUT&Tag is a new method for studying protein-DNA interactions. Compared with traditional ChIP-seq, it has the following advantages: time-saving and high efficiency, small amount of cells required, low background signal, good repeatability, etc. It can even be used for single-cell sequencing. CUT&Tag is a powerful tool for studying the interaction between proteins and chromatin DNA in vivo, and is of great significance for research on gene regulation, epigenetics, etc.
Version 2.0 of ChiTag® uses pAG-Tn5 to replace the original pA-Tn5, which improves efficiency and removes the restrictions of antibody selection

Usage

Protein-dna interaction can replace ChIP-Seq and other methods. It is especially suitable for efficient epigenome analysis of small samples and single cells.

Quality Control

After multiple column purifications, only a clear single target band was visible in SDS-PAGE gel detection, and no host DNA residue was detected by PCR method, and no endonuclease or exonuclease contamination was found.

Storage

It can be stored at -20°C for 3 months and at -70°C for long-term storage.

Components

Component	M058-YH01-01A Size: 12 rxns
NovoNGS® ChiTag® pAG-Transposase (6.5pmol/μl)	12μl
Annealing Buffer	500μl

Applications

1. Translocation complex Construction (ChiTag® Translocation)
A. lyophilized powder dissolved
The freeze-dried powder of single-chain ME, single-chain adapter1 and single-chain adapter2 were centrifuged at high speed for 1min and added to the Annealing Buffer to dissolve, respectively, with the dissolution concentration of 100μM. Vortex mixing to ensure the joint fully dissolved.
B. Prepare two kinds of joints
adapter1 and adapter2 were mixed in equal volume and ME and Adapter2 were mixed in equal volume, and then cooled and annealed at a low speed by PCR. PCR was set to 95℃ for 3min; 95℃ to 25℃ for 45min; 4℃Hold.
C. Complex preparation
The hybrid joint is obtained by mixing joint 1 and joint 2 in equal volume. The enzyme and the joint were mixed evenly and incubated at room temperature for 1h. Usually the molar ratio of the enzyme mixed with the joint is 1:1, and the incubation ratio of the joint can be increased as needed.
If the ratio of enzyme to joint mole is 1:1, the configuration is as follows:
Double link head mixture (50pmol/μl),,1μl; NovoNGS® ChiTag® 2.0 Transposase (6.5pmol/μl), 7.7μl
2. CUT&Tag experimental test
Cut&Tag experiment was conducted with 80,000 cells, and goat antibody was used as the secondary antibody. As shown in the figure below, the library yield obtained was more than that of pA-Tn5. The use of pA-Tn5 instead of PA-TN5 could reduce the requirement for secondary antibody in the experiment.

Legend: Lane B: negative control (without primary antibody); Lane C: pA-Tn5 CUT&Tag amplification product; Lane D: pAG-Tn5 CUT&Tag amplification product; MK: DNA Maker.

For Research or Manufacturing Use Only, Not for Use in Diagnostic or Therapeutic Procedures.

Novoprotein is a trademark of Novoprotein Scientific Inc. visit our trademark information page: TradeMark.

All other trademarks are the property of their respective owners.

Purchasing：

+86 400-600-0940

Product：

product@novoprotein.com

Service：

service@novoprotein.com.cn

Support：

support@novoprotein.com.cn

Resource Download