Description
LZCap™ AG(3'Acm) is a Cap1 analog, can be used as the capping agent for producing mRNAs in an "one-pot"process. Under the action of T7 polymerase, mRNA with 5' end Cap 1 structure was generated by cotranscription using LZCap™ AG(3'Acm), NTPs, and template DNA. The capped mRNA could be directly translated and expressed in cells and in vivo. It is widely used in the fields of in vitro transcription, gene editing, vaccine development, tumor CAR-T therapy, protein substitution therapy and regenerative medicine.
LZCap™ AG(3'Acm) requires the T7 promoter with AG as the starting sequence. LZCap™ AG(3'Acm) can provide>97%capped mRNA, and up to 100-200 ug capped mRNA can be generated with 1 ug DNA template per standard reaction
Specifications
|
Cat.No. |
10684ES60/10684ES80 |
|
Size |
100 uL/1mL |
|
Molecular Formula |
C35H48N16O24P4（Free acid） |
|
Molecular Weight |
1200.75（Free acid） |
|
Concentration |
100 mM |
|
Purity |
HPLC ≥90% |
|
Salt type |
NH4+ |
|
Structure |
|
Components
|
Name |
10684ES60 |
10684ES80 |
|
LZCap™ AG(3'Acm) GMP-grade (100 mM) |
100 μL |
1 mL |
Storage
This product should be stored at -25~-15℃ for 2 years
Instructions
LZCap™ DNA Template Design
LZCap™ AG(3'Acm) is suitable for AG-initiated sequences. As shown in the figure below, the T7 promoter (underlined) followed by the AG sequence can effectively initiate transcription.
1. Thaw components required for the experiment on ice.
2.Refer to the following reaction system to configure the transcription system at room temperature.
|
Component |
Volume(μL) |
Final concentration |
|
ATP(100mM) |
1 |
5mM |
|
UTP(100mM) |
1 |
5mM |
|
CTP(100mM) |
1 |
5mM |
|
GTP(100mM) |
1 |
5mM |
|
LZCap™ AG(3'Acm) (100mM) |
0.8 |
4mM |
|
10×Transcription Buffer |
2 |
1× |
|
Recombinant RNase Inhibitor(40U/μL) |
0.5 |
1U/μL |
|
Pyrophosphatase(0.1U/μL) |
0.4 |
0.002U/μL |
|
T7 RNA polymers(250U/μL) |
0.64 |
8U/μL |
|
Linear DNA+RNase Free Water |
11.66 |
1 μg |
|
Final volume |
20μL |
|
*Modified N-Me-pUTP can be used in place of wild-type UTP. The modified N-Me-pUTP reduces the immunogenicity of mRNA.
3.Mix the prepared reaction solution, centrifuge briefly, and incubate at 37C for 2-3 hours. If the transcript
length is less than 100nt, increase the reaction time to 4-8 h.
Notes
1.LZCap™ AG(3'Acm) is suitable for T7 promoter transcription vector with 5 'AG 3' initiated sequences, which needs to be considered when constructing the vector.
2. The reagents, consumables and containers used in the experiment are free of RNase contamination.
3.It is recommended to use a linearized DNA template for transcription.
4.When modified nucleotides were used in place of wild-type nucleotides, the final concentration of the reaction was unchanged.
5.If the PCR product is used as the transcription initiation DNA template, the amount of DNA template can be reduced by half.
6.This product is for research use only.
7.Please operate with lab coats and disposable gloves, for your safety.
Documents:
Safety Data Sheet
10684_MSDS_HB250613.pdf
Manuals
10684_Manual_Ver.EN20250613.pdf
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The product is for research purposes only and is not intended for therapeutic or diagnostic use in humans or animals. Products and content are protected by patents, trademarks, and copyrights owned by Yeasen Biotechnology. Trademark symbols indicate the country of origin, not necessarily registration in all regions.
Certain applications may require additional third-party intellectual property rights.
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