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Mmessage mmachine high yield capped rna kit

Manufactured by Thermo Fisher Scientific
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The MMESSAGE mMACHINE high yield capped RNA kit is a lab equipment product designed for the in vitro transcription of capped RNA. The kit provides the necessary components to generate high yields of capped RNA from DNA templates.

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Lab products found in correlation

Axiovert microscope, by Zeiss (2 mentions) Axiocam mr camera, by Zeiss (1 mentions) Lumar v12, by Zeiss (1 mentions) Pgem t easy, by Promega (1 mentions)

Close spelling variants of this product

T7 mMESSAGE mMACHINE™ High Yield Capped RNA Transcription Kit MMESSAGE mMACHINE Kit High Yield Capped RNA Transcription Kit MMESSAGE mMACHINE High Yield Capped RNA Transcription kit SP6 mMessage mMachine High Yield Capped RNA Transcription Kit MMessage mMachine kit MMessage mMachine

5 protocols using mmessage mmachine high yield capped rna kit

1

Sodium Channel Expression in Oocytes

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The procedures for oocyte preparation, cRNA synthesis and injection were identical to those described previously [31 (link)]. cRNA was prepared by in vitro transcription with T7 polymerase using the mMESSAGE mMACHINE high yield capped RNA kit (Ambion, Austin, TX). To enhance the expression of BgNav2-1 and BgNav1-1a and the mutant channels, their cRNAs (2-5 ng/oocyte) were co-injected into oocytes with tipE cRNA from Drosophila melanogaster in a 1:1 ratio [32 (link), 33 (link)]
Zhorov B.S., Du Y., Song W., Luo N., Gordon D., Gurevitz M, & Dong K. (2021). Mapping the interaction surface of scorpion β-toxins with an insect sodium channel. The Biochemical journal, 478(14), 2843-2869.
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2

Xenopus Oocyte Sodium Channel Expression

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Ovaries from oocyte-positive female Xenopus laevis were purchased from Xenopus 1 (Dexter, MI). The procedures for oocyte preparation, cRNA synthesis and injection were identical to those described previously (Tan et al., 2002a (link)). cRNA was prepared by in vitro transcription with T7 polymerase using the mMESSAGE mMACHINE® high yield capped RNA kit (Ambion, Austin, TX). For robust expression of the BgNav1-1a sodium channels, the BgNav1-1a cRNAwas co-injected into oocytes with Drosophila melanogaster tipE cRNA (1:1 M ratio), which enhances the expression of insect sodium channels (Feng et al., 1995 (link); Warmke et al., 1997 (link)), but does not alter the sensitivity of cockroach sodium channels to pyrethroids (Data not shown).
Chen M., Du Y., Nomura Y., Zhu G., Zhorov B.S, & Dong K. (2017). Mutations of two acidic residues at the cytoplasmic end of segment IIIS6 of an insect sodium channel have distinct effects on pyrethroid resistance. Insect biochemistry and molecular biology, 82, 1-10.
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3

Heterologous Expression of AaNa(v)1-1 in Xenopus Oocytes

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AaNav1-1 and mutants were expressed in the Xenopus oocytes, Ovaries from oocyte-positive female Xenopus laevis purchased from Xenopus 1 (Dexter, MI). The procedures for oocyte preparation, cRNA synthesis and injection were identical to those described previously [35 (link)]. cRNA was prepared by in vitro transcription with T7 polymerase using the mMESSAGE mMACHINE high yield capped RNA kit (Ambion, Austin, TX). To enhance expression of AaNav1-1 and mutant channels, their cRNAs were co-injected into oocytes with Ae. aegypti tipE cRNA in the 1:1 molar ratio [36 (link), 37 (link)].
Chen M., Du Y., Wu S., Nomura Y., Zhu G., Zhorov B.S, & Dong K. (2019). Molecular evidence of sequential evolution of DDT- and pyrethroid-resistant sodium channel in Aedes aegypti. PLoS Neglected Tropical Diseases, 13(6), e0007432.
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4

Developmental Gene Expression in Zebrafish

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Total RNA was extracted from zebrafish embryos at various stages of development to amplify a RT-PCR fragment of the zebrafish orthologues for lim3 (20) , slc35d3 and pax2a (primers available upon request). These fragments were cloned into pGEM-T-easy (Promega) for making antisense probes by linearization of the plasmid and subjected to in vitro transcription using the mMessage mMachine High Yield Capped RNA kit (Ambion). Developmental staging of injected zebrafish was determined by somite number and embryos were fixed in 4% paraformaldehyde.
Assays for RNA expression using WISH were performed as described (21) . Embryos were visualized with a Zeiss Lumar V12 or a Zeiss Axiovert microscope and images were captured with a Zeiss Axiocam MR camera using AxioVision software.
Buyse G., Di Michele M., Wijgaerts A., Louwette S., Wittevrongel C., Thys C., Downes K., Ceulemans B., Van Esch H., Van Geet C, & Freson K. (2020). Unravelling the disease mechanism for TSPYL1 deficiency. Human molecular genetics, 29(20).
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5

Robust Expression of Insect Sodium Channels in Oocytes

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The procedures for oocyte preparation, cRNA synthesis and injection were identical to those described previously (Tan et al., 2002 (link)). cRNA was prepared by in vitro transcription with T7 polymerase using the mMESSAGE mMACHINE® high yield capped RNA kit (Ambion, Austin, TX). For robust expression of the BgNav1-1a sodium channels, BgNav1-1a cRNA was coinjected into oocytes with Drosophila melanogaster tipE cRNA (1:1 molar ratio) which enhances the expression of insect sodium channels in oocytes (Feng et al., 1995 (link); Warmke et al., 1997 (link)).
Chen M., Du Y., Nomura Y., Zhu G., Zhorov B.S, & Dong K. (2016). Alanine to valine substitutions in the pore helix IIIP1 and linker-helix IIIL45 confer cockroach sodium channel resistance to DDT and pyrethroids. Neurotoxicology, 60, 197-206.
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