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Ambion t7 megashortscript kit

Manufactured by Thermo Fisher Scientific
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The Ambion T7 MEGAshortscript kit is a laboratory product designed for the in vitro transcription of small RNA molecules. It provides a rapid and efficient method for the synthesis of small RNAs, such as siRNAs and microRNAs, from DNA templates.

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Lab products found in correlation

Ambion megaclear kit, by Thermo Fisher Scientific (2 mentions) Column pcr purification kit, by Qiagen (2 mentions) Nanodrop instrument, by Thermo Fisher Scientific (2 mentions) Agilent bioanalyzer, by Agilent Technologies (2 mentions) Hiv 1 rt, by Worthington (2 mentions) Superase in, by Thermo Fisher Scientific (2 mentions) Proteinase k, by Thermo Fisher Scientific (2 mentions) Gel loading buffer 2, by Thermo Fisher Scientific (1 mentions) T4 polynucleotide kinase, by Thermo Fisher Scientific (1 mentions) Dna oligonucleotide, by Integrated DNA Technologies (1 mentions) γ 32p atp, by PerkinElmer (1 mentions) γ 33p atp, by PerkinElmer (1 mentions) Dna and rna oligonucleotides, by Integrated DNA Technologies (1 mentions) α 33p gtp, by PerkinElmer (1 mentions)

4 protocols using ambion t7 megashortscript kit

1

Efficient sgRNA Generation via T7 Amplification

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T7 promoter was added to sgRNA template via PCR amplification using the Mmp21 primers (Supplementary Table 12): p(Ile226Thr)-T7-forward (near orthologous p.(Ile226Thr)) and p(Ala321Pro)-T7-forward (near orthologous p.(Ala321Pro)) and common reverse primer containing the stemloop,. The PCR product was purified with a column PCR purification kit (Qiagen). In vitro transcription was then performed with an Ambion T7 MEGAshortscript kit (Life Technologies) and RNA was recovered using an Ambion MEGAclear kit (Life Technologies). The concentration of RNA was measure by a NanoDrop instrument (Thermo Scientific) and Agilent Bioanalyzer (Agilent Technologies, Inc).
Guimier A., Gabriel G.C., Bajolle F., Tsang M., Liu H., Noll A., Schwartz M., El Malti R., Smith L.D., Klena N.T., Jimenez G., Miller N.A., Oufadem M., Moreau de Bellaing A., Yagi H., Saunders C.J., Baker C.N., Di Filippo S., Peterson K.A., Thiffault I., Bole-Feysot C., Cooley L.D., Farrow E.G., Masson C., Schoen P., Deleuze J.F., Nitschké P., Lyonnet S., de Pontual L., Murray S.A., Bonnet D., Kingsmore S.F., Amiel J., Bouvagnet P., Lo C.W, & Gordon C.T. (2015). MMP21 is mutated in human heterotaxy and is required for normal left-right asymmetry in vertebrates. Nature genetics, 47(11), 1260-1263.
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2

Efficient sgRNA Generation via T7 Amplification

Check if the same lab product or an alternative is used in the 5 most similar protocols
T7 promoter was added to sgRNA template via PCR amplification using the Mmp21 primers (Supplementary Table 12): p(Ile226Thr)-T7-forward (near orthologous p.(Ile226Thr)) and p(Ala321Pro)-T7-forward (near orthologous p.(Ala321Pro)) and common reverse primer containing the stemloop,. The PCR product was purified with a column PCR purification kit (Qiagen). In vitro transcription was then performed with an Ambion T7 MEGAshortscript kit (Life Technologies) and RNA was recovered using an Ambion MEGAclear kit (Life Technologies). The concentration of RNA was measure by a NanoDrop instrument (Thermo Scientific) and Agilent Bioanalyzer (Agilent Technologies, Inc).
Guimier A., Gabriel G.C., Bajolle F., Tsang M., Liu H., Noll A., Schwartz M., El Malti R., Smith L.D., Klena N.T., Jimenez G., Miller N.A., Oufadem M., Moreau de Bellaing A., Yagi H., Saunders C.J., Baker C.N., Di Filippo S., Peterson K.A., Thiffault I., Bole-Feysot C., Cooley L.D., Farrow E.G., Masson C., Schoen P., Deleuze J.F., Nitschké P., Lyonnet S., de Pontual L., Murray S.A., Bonnet D., Kingsmore S.F., Amiel J., Bouvagnet P., Lo C.W, & Gordon C.T. (2015). MMP21 is mutated in human heterotaxy and is required for normal left-right asymmetry in vertebrates. Nature genetics, 47(11), 1260-1263.
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3

Identification and Characterization of HIV-1 and SIV RNA Elements

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DNA oligonucleotides and pIDTSMART vectors were purchased from Integrated DNA Technologies (IDT) (Coralville, IA, USA). [γ-32P]ATP was obtained from PerkinElmer (Shelton, CT, USA). HIV-1 RT was purchased from Worthington (Lakewood, NJ, USA). T4 polynucleotide kinase, proteinase K, SUPERaseIN, and Gel Loading Buffer II were bought from Life Technologies (Foster City, CA, USA). E271 loading dye base was obtained from AMRESCO LLC (Solon, OH, USA). The Ambion MEGAshortscript T7 kit was purchased from Life Technologies. The sequences of the HIV-1 acceptor RNA and (−) SSDNA, as well as the TAR and Psi RNAs were derived from HIV-1 NL4-3 (GenBank Accession no. AF324493) [94 (link)]. The corresponding SIV NAs were derived from SIVmac239 (GenBank Accession no. M33262) [12 (link), 13 (link)], which was obtained from Dr. Ronald Desrosiers through the AIDS Reagent Program, Division of AIDS, NIAID, NIH.
Post K., Olson E.D., Naufer M.N., Gorelick R.J., Rouzina I., Williams M.C., Musier-Forsyth K, & Levin J.G. (2016). Mechanistic differences between HIV-1 and SIV nucleocapsid proteins and cross-species HIV-1 genomic RNA recognition. Retrovirology, 13, 89.
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4

Preparation of Radiolabeled HIV-1 Nucleic Acid Substrates

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DNA and RNA oligonucleotides were obtained from Integrated DNA Technologies (IDT) (Coralville, IA). T4 polynucleotide kinase, proteinase K, SUPERaseIN, and Gel Loading Buffer II were purchased from Applied Biosystems (now Life Technologies) (Foster City, CA). [γ-33P]ATP, [α-33P]dCTP, and [α-33P]GTP were purchased from PerkinElmer (Shelton, CT). Purified tRNALys3 from human placenta was obtained from Bio S&T (Lachine, Québec, Canada). RNA transcripts were synthesized by using the Ambion MEGAshortscript T7 kit (Life Technologies). HIV-1 RT was obtained from Worthington (Lakewood, NJ). The SP2 peptide (FLGKIWPSHKGRPGNF) was purchased from GenScript (Piscataway, NJ). The sequences of the viral nucleic acid substrates and the RNA 200 and RNA 60 templates were derived from HIV-1 NL4-3 (GenBank Accession no. AF324493) (Adachi et al., 1986 (link)). The RNA 105 template was derived from the HIV-1 HXB2 sequence (GenBank Accession no. K03455), as described previously (Hargittai et al., 2001 (link)).
Wu T., Gorelick R.J, & Levin J.G. (2014). Selection of fully processed HIV-1 nucleocapsid protein is required for optimal nucleic acid chaperone activity in reverse transcription. Virus research, 193, 52-64.
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