T7 rna polymerase

Manufactured by Enzynomics

T7 RNA polymerase is an enzyme used in molecular biology and biotechnology. It is responsible for the transcription of DNA into RNA, specifically targeting the T7 promoter sequence. This enzyme is essential for various applications, such as in vitro RNA synthesis and gene expression studies.

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Lab products found in correlation

Hiscribe t7 high yield rna synthesis kit, by New England Biolabs (1 mentions) Monarch rna cleanup kit, by New England Biolabs (1 mentions) Dnase 1, by New England Biolabs (1 mentions) Dnase 1 reaction buffer, by New England Biolabs (1 mentions) Nebuffer r2, by New England Biolabs (1 mentions) Dnase 1, by Enzynomics (1 mentions) Uracil dna glycosylase, by New England Biolabs (1 mentions) Ultrapure dnase rnase free distilled water, by Bioneer (1 mentions) Dfhbi 1t, by Merck Group (1 mentions) Dfhbi, by Merck Group (1 mentions)

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T7 polymerase (1 citations)

3 protocols using t7 rna polymerase

CRISPR-Cas13a-based Biomolecular Assay

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All oligonucleotides (Table S1) used in this study were purchased from Bionics (Seoul, Republic of Korea) and Integrated DNA Technologies (Coralville, IA, USA). T7 RNA polymerase, T7 RNA polymerase buffer, and RNase inhibitors were obtained from Enzynomics (Daejeon, Republic of Korea). The HiScribe T7 High Yield RNA synthesis kit, Monarch RNA cleanup kit, DNase I, DNase I reaction buffer, NEBuffer™ r2.1, and rNTP were purchased from New England Biolabs (Ipswich, MA, USA). Plates with 384 black wells were purchased from SPL Life Sciences (Pocheon, Republic of Korea). To1-3PEG-Biotin Fluorophore (To1-biotin) was obtained from Applied Biological Materials (Richmond, BC, Canada). LbuCas13a was purchased from SignalChem Diagnostics (St. Louis, MO, USA).

Kim Y., Nam D., Lee E.S., Kim S., Cha B.S, & Park K.S. (2024). Aptamer-Based Switching System for Communication of Non-Interacting Proteins. Biosensors, 14(1), 47.

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Preparation of HIV-1 Transcripts and Labeled Oligonucleotides

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Fragments of the HIV-1 5′ LTR, gag, and 3′ LTR were transcribed in vitro using T7 RNA polymerase (Enzynomics). The transcripts were purified using denaturing polyacrylamide gel electrophoresis (PAGE; 5% polyacrylamide, 8 M urea). Synthetic DNA and RNA oligonucleotides were 5′-end labeled with ³²P using T4 polynucleotide kinase and [γ-³²P]ATP. The nucleotides were precipitated by the addition of 3 M ammonium acetate and 100% (vol/vol) ethanol. The nucleotides pellets were washed with 75% ethanol, dried and resuspended in 10 mM Tris-Cl (pH 7.8) containing 1 mM EDTA. The synthetic dsDNAs and dsRNAs (Supplementary Table 1) were prepared by annealing the oligonucleotides.

Ryoo J., Choi J., Oh C., Kim S., Seo M., Kim S., Seo D., Kim J., White T.E., Brandariz-Nunez A., Diaz-Griffero F., Yun C.H., Hollenbaugh J.A., Kim B., Baek D, & Ahn K. (2014). The ribonuclease activity of SAMHD1 is required for HIV-1 restriction. Nature medicine, 20(8), 936-941.

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Oligonucleotide Synthesis and Purification

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Bioneer Co. (Daejeon, South Korea) synthesized and purified the oligonucleotides (Table S1†) used in this work via polyacrylamide gel electrophoresis (PAGE), except for the hairpin substrate probe (HP), which underwent purification by HPLC. DNase I, Escherichia coli (E. coli) RNase H, ribonucleoside triphosphate (rNTP) set, exonuclease I (Exo I), exonuclease III (Exo III), and T7 RNA polymerase were obtained from Enzynomics (Daejeon, South Korea). Lambda exonuclease (λ Exo), Eco RI, and Uracil DNA glycosylase (UDG) were supplied by New England Biolabs Inc. (Beverly, MA, USA). DFHBI and DFHBI-1T were acquired from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure DNase/RNase-free distilled water was supplied by Bioneer Co. and used for all assays. Other reagents used in this work were of analytical grade and used without any additional purification.

Lee J., Kim H., Li Y., Lee S, & Park H.G. (2024). An ultrasensitive label-free RNase H assay based on in vitro transcription of fluorogenic light-up aptamer. Nanoscale Advances, 6(7), 1926-1931.

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