LZCap GG (3'Acm) is a cap1 analog. The product is used for transcription with the initial sequence of 5'GG3', and the natural cap1 structure is produced by Cap1 transcription capping. US patent approved. Components Cat No. Name Size 10686 LZCap GG (3'Acm) (50mM) 100 μL 1 mL Product Details Molecular Formula C35H48N16O25P4 (Free acid) Molecular weight 1216.19 (Free acid) Concentration 50± 3 mM Purity HPLC ≥95% LZCap DNA Template Design LZCap GG(3'Acm) is suitable for GG-initiated sequences. As shown in the figure below, the SP6 promoter (underlined) followed by the GG sequence can effectively initiate transcription. Protocols 1． Thaw components required for the experiment on ice. 2． Add RNase free water and NTPs to reaction tube. 3． Heat LZCap GG(3'Acm) aliquot for 15 min at 60°C. Cool at room temperature for 5 min. 4． Immediately add LZCap GG(3'Acm) to tube and vortex to mix. Spin briefly to collect liquid. 5． Add 10x Transcription Buffer and vortex. 6． Add DNA template and SP6 Enzyme Mix. 7． Mix the prepared reaction solution, spin briefly, and incubate at 37°C for 2-3 hours. If the transcript length is less than 100nt, increase the reaction time to 4-8 h. Component Volume (μL) Final concentration RNase Free Water Up to 20μL / ATP(100mM) 1 5mM UTP(100mM) 1 5mM CTP(100mM) 1 5mM GTP(100mM) 0.2 1mM LZCap GG(3'Acm) (50mM) 1.6 4mM 10×Transcription Buffer 2 1 × Linear DNA 1μg 50 ng/μL SP6 Enzyme Mix 2 / Final Volume 20μL Notes: 1) The SP6 enzyme mix we recommended to used is MEGAscript® Kit（AM1330）. 2) LZCap GG(3'Acm) is suitable for SP6 promoter transcription vector with 5 ‘GG 3’initiated sequences, which needs to be considered when constructing the vector. 3) The reagents, consumables and containers used in the experiment are free of RNase contamination. 4) It is recommended to use a linearized DNA template for transcription. 5) When modified nucleotides were used in place of wild-type nucleotides, the final concentration of the reaction was unchanged. 6) Modified N1-Me-pUTP can be used in place of wild-type UTP. The modified N1-Me-pUTP reduces the immunogenicity of mRNA. Henovcom can also provide modified nucleotide N1-Me-pUTP (Cat. No.: HN1002). 7) If the PCR product is used as the transcription initiation DNA template, the amount of DNA template can be reduced by half. Precautions For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product. For research use only. Please contact us for more details about the LZCap licensing.
