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Megaclear transcription reaction purification kit

Manufactured by Thermo Fisher Scientific
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The MEGAclear transcription reaction purification kit is a tool for the purification of in vitro transcribed RNA. It is designed to remove unincorporated nucleotides, salts, and other reaction components from the final RNA product.

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Lab products found in correlation

Megashortscript in vitro transcription kit, by Thermo Fisher Scientific (3 mentions) Small rna analysis kit, by Agilent Technologies (3 mentions) Agilent bioanalyzer 2100 system, by Agilent Technologies (3 mentions) Kod hot start polymerase, by Merck Group (3 mentions)

3 protocols using megaclear transcription reaction purification kit

1

In Vitro Transcription of RNA Templates

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DNA templates for in vitro transcription of scrambled RNA, 5S rRNA, and RNA5SP141 were generated by PCR with the primers listed in Supplementary Fig. 3b using KOD Hot Start Polymerase (EMD Millipore). DNA templates for in vitro transcription for control rabies virus leader sequence (RABVLe) and rabies virus internal sequence (RABVINT) were generated by annealing sense and antisense oligonucleotides as previously described20 (link). RNA was synthesized using the MEGAshortscript in vitro transcription kit (Ambion) and purified using the MEGAclear transcription reaction purification kit (Ambion). RNA integrity was evaluated with the Agilent 2100 Bioanalyzer System using the Small RNA Analysis Kit (Agilent).
Chiang J.J., Sparrer K.M., van Gent M., Lässig C., Huang T., Osterrieder N., Hopfner K.P, & Gack M.U. (2017). Viral unmasking of cellular 5S rRNA pseudogene transcripts induces RIG-I mediated immunity. Nature immunology, 19(1), 53-62.
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2

RNA Synthesis and Purification Protocol

Cited 1 time
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DNA templates for the in vitro transcription of scrambled RNA and RNA5SP141 were generated by PCR as described previously (12 (link)) using KOD Hot Start Polymerase (EMD Millipore). DNA templates for the in vitro transcription of rabies virus leader sequence (RABVle) were generated by annealing sense and antisense oligonucleotides as previously described (54 (link)). RNA was synthesized using the MEGAshortscript in vitro transcription kit (Ambion) and purified using the MEGAclear transcription reaction purification kit (Ambion). RNA integrity was evaluated with the Agilent 2100 Bioanalyzer System using the Small RNA Analysis Kit (Agilent).
Naesens L., Muppala S., Acharya D., Nemegeer J., Bogaert D., Lee J.H., Staes K., Debacker V., De Bleser P., De Bruyne M., De Baere E., van Gent M., Liu G., Lambrecht B.N., Staal J., Kerre T., Beyaert R., Maelfait J., Tavernier S.J., Gack M.U, & Haerynck F. (2022). GTF3A mutations predispose to herpes simplex encephalitis by disrupting biogenesis of the host-derived RIG-I ligand RNA5SP141. Science immunology, 7(77), eabq4531.
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3

In Vitro Transcription of RNA Templates

Check if the same lab product or an alternative is used in the 5 most similar protocols
DNA templates for in vitro transcription of scrambled RNA, 5S rRNA, and RNA5SP141 were generated by PCR with the primers listed in Supplementary Fig. 3b using KOD Hot Start Polymerase (EMD Millipore). DNA templates for in vitro transcription for control rabies virus leader sequence (RABVLe) and rabies virus internal sequence (RABVINT) were generated by annealing sense and antisense oligonucleotides as previously described20 (link). RNA was synthesized using the MEGAshortscript in vitro transcription kit (Ambion) and purified using the MEGAclear transcription reaction purification kit (Ambion). RNA integrity was evaluated with the Agilent 2100 Bioanalyzer System using the Small RNA Analysis Kit (Agilent).
Chiang J.J., Sparrer K.M., van Gent M., Lässig C., Huang T., Osterrieder N., Hopfner K.P, & Gack M.U. (2017). Viral unmasking of cellular 5S rRNA pseudogene transcripts induces RIG-I mediated immunity. Nature immunology, 19(1), 53-62.
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