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unmethylated gpppg (New England Biolabs)

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New England Biolabs unmethylated gpppg
Unmethylated Gpppg, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rna+cap+analog/G(5)ppp(5)G+RNA+Cap+Struc+Analog/pm42103131-88-26-28
Average 95 stars, based on 187 article reviews

unmethylated gpppg - by Bioz Stars, 2026-09

95/100 stars

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Incubation:

Article Title: High-resolution structures of the SARS-CoV-2 N7-methyltransferase inform therapeutic development
Article Snippet: The MTase activity was measured using the MTase-Glo Methyltransferase bioluminescence assay (Promega) 19 following the manufacturer’s instructions. .. The reaction mix containing 20 mM Tris pH 8.0, 50 mM NaCl, 1 mM EDTA, 3 mM MgCl 2 , 0.1 mg ml –1 BSA, 1 mM DTT, 20 μM protein (nsp14/10, MTase domain or TEL-MTase), 20 μM SAM and 0.15 mM G(5′)ppp(5′)A RNA cap analog (NEB, S1406S) were incubated for 1 h at room temperature. ..

Article Title: Despite the odds: formation of the SARS-CoV-2 methylation complex.
Article Snippet: .. Briefly, the methyltransferase reaction was incubated for 20 min at RT in 8 μl reaction volume with 100 nM nsp10 / 16 / 14wt or mutated heterotrimer, 1 μM Ultrapure SAM (from the Cisbio’s kit), 187.5 μM RNA cap analog (GpppA or GpppG, New England Biolabs S1406S and S1407S, respectively) or 18.75 μM Cap 0 RNA oligo (TriLink, produced on request) in reaction buffer consisting of 20 mM Tris–HCl pH 7.4, 150 mM NaCl and 0.5 mM DTT. .. The reaction was quenched by adding 2 μl of 5M NaCl followed by the addition of 2 μl Detection Buffer 1 (from the Cisbio’s kit) to the reaction mixture.

Article Title: Despite the odds: formation of the SARS-CoV-2 methylation complex
Article Snippet: .. Briefly, the methyltransferase reaction was incubated for 20 min at RT in 8 μl reaction volume with 100 nM nsp10/16/14wt or mutated heterotrimer, 1 μM Ultrapure SAM (from the Cisbio's kit), 187.5 μM RNA cap analog (GpppA or GpppG, New England Biolabs S1406S and S1407S, respectively) or 18.75 μM Cap 0 RNA oligo (TriLink, produced on request) in reaction buffer consisting of 20 mM Tris–HCl pH 7.4, 150 mM NaCl and 0.5 mM DTT. .. The reaction was quenched by adding 2 μl of 5M NaCl followed by the addition of 2 μl Detection Buffer 1 (from the Cisbio's kit) to the reaction mixture.

Ligation:

Article Title: Evidence for a Genetic and Physical Interaction between Nonstructural Proteins NS1 and NS4B That Modulates Replication of West Nile Virus
Article Snippet: .. After ligation, phenol-chloroform extraction, and ethanol precipitation, DNA-dependent RNA transcription was performed in vitro using the Ampliscribe T7 RNA polymerase transcription kit (Epicentre) with an m7G(5=)ppp(5=)A RNA cap analog (New England BioLabs). .. In vitro transcribed SINV or WNV RNA was electroporated to BHK21-15 cells using a GenePulser Xcell electroporator (Bio-Rad) at 850 V, 25 mF, and infinite to generate a P0 virus stock.

Article Title: Evidence for a Genetic and Physical Interaction between Nonstructural Proteins NS1 and NS4B That Modulates Replication of West Nile Virus
Article Snippet: .. After ligation, phenol-chloroform extraction, and ethanol precipitation, DNA-dependent RNA transcription was performed in vitro using the Ampliscribe T7 RNA polymerase transcription kit (Epicentre) with an m 7 G(5′)ppp(5′)A RNA cap analog (New England BioLabs). .. In vitro transcribed SINV or WNV RNA was electroporated to BHK21-15 cells using a GenePulser Xcell electroporator (Bio-Rad) at 850 V, 25 mF, and infinite Ω to generate a P0 virus stock.

Extraction:

Article Title: Evidence for a Genetic and Physical Interaction between Nonstructural Proteins NS1 and NS4B That Modulates Replication of West Nile Virus
Article Snippet: .. After ligation, phenol-chloroform extraction, and ethanol precipitation, DNA-dependent RNA transcription was performed in vitro using the Ampliscribe T7 RNA polymerase transcription kit (Epicentre) with an m 7 G(5′)ppp(5′)A RNA cap analog (New England BioLabs). .. In vitro transcribed SINV or WNV RNA was electroporated to BHK21-15 cells using a GenePulser Xcell electroporator (Bio-Rad) at 850 V, 25 mF, and infinite Ω to generate a P0 virus stock.

Ethanol Precipitation:

Article Title: Evidence for a Genetic and Physical Interaction between Nonstructural Proteins NS1 and NS4B That Modulates Replication of West Nile Virus
Article Snippet: .. After ligation, phenol-chloroform extraction, and ethanol precipitation, DNA-dependent RNA transcription was performed in vitro using the Ampliscribe T7 RNA polymerase transcription kit (Epicentre) with an m 7 G(5′)ppp(5′)A RNA cap analog (New England BioLabs). .. In vitro transcribed SINV or WNV RNA was electroporated to BHK21-15 cells using a GenePulser Xcell electroporator (Bio-Rad) at 850 V, 25 mF, and infinite Ω to generate a P0 virus stock.

In Vitro:

Article Title: Evidence for a Genetic and Physical Interaction between Nonstructural Proteins NS1 and NS4B That Modulates Replication of West Nile Virus
Article Snippet: .. After ligation, phenol-chloroform extraction, and ethanol precipitation, DNA-dependent RNA transcription was performed in vitro using the Ampliscribe T7 RNA polymerase transcription kit (Epicentre) with an m 7 G(5′)ppp(5′)A RNA cap analog (New England BioLabs). .. In vitro transcribed SINV or WNV RNA was electroporated to BHK21-15 cells using a GenePulser Xcell electroporator (Bio-Rad) at 850 V, 25 mF, and infinite Ω to generate a P0 virus stock.

Produced:

Concentration Assay:

Article Title: The Mammalian Cap-Specific m 6 Am RNA Methyltransferase PCIF1 Regulates Transcript Levels in Mouse Tissues.
Article Snippet: .. As for TrypPcif1, the protocol is same as dPcif1, except that the concentration of protein is 30 mM and the concentration of RNA cap analog is 300 mM [m7G(5)ppp(5)G (NEB, Cat. No. S1404S) or m7G(5)ppp(5)A (NEB, Cat. No. S1405S) or m7G(5)ppp(5)(20OMeA)pG (Trilink, Cat. No. N-7413)]. ..

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$(A) A-cap / stemloop IRES reporter RNA carrying the negative control “scramble 1” sequence was mixed into a polysome lysate made from HEK293T cells for five minutes on ice and loaded on a sucrose gradient. (B) The sucrose gradient was fractionated and cDNA was prepared from groups of fractions as shown (numbered 1 -6). qRT-PCR was performed to quantify the exogenous “scramble 1” RNA across the fractions, as compared to endogenous Gapdh mRNA. The “scramble 1” transcript migrated into polysomal fractions 4-6, even though it is not translated .$

Journal: bioRxiv

Article Title: Evaluating the reliability of tools for mRNA annotation and IRES studies

doi: 10.64898/2026.03.29.707813

Figure Lengend Snippet: (A) A-cap / stemloop IRES reporter RNA carrying the negative control “scramble 1” sequence was mixed into a polysome lysate made from HEK293T cells for five minutes on ice and loaded on a sucrose gradient. (B) The sucrose gradient was fractionated and cDNA was prepared from groups of fractions as shown (numbered 1 -6). qRT-PCR was performed to quantify the exogenous “scramble 1” RNA across the fractions, as compared to endogenous Gapdh mRNA. The “scramble 1” transcript migrated into polysomal fractions 4-6, even though it is not translated .

Article Snippet: The ARCA m 7 G-cap analog (New England Biolabs; S1411) was used at a 4:1 ratio to GFP for transcription of T7-nluc constructs, while an A-cap analog (New England Biolabs S1406), was used to generate T7HP-nluc IRES reporter RNAs.

Techniques: Negative Control, Sequencing, Quantitative RT-PCR