Sp6 mmessage mmachine high yield capped rna transcription kit

Manufactured by Thermo Fisher Scientific

The SP6 mMessage mMachine High Yield Capped RNA Transcription Kit is a laboratory equipment product that enables the in vitro synthesis of capped and polyadenylated RNA transcripts from SP6 RNA polymerase promoter-driven DNA templates. The kit includes reagents and enzymes necessary for the transcription reaction.

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Lab products found in correlation

Femtojet, by Eppendorf (1 mentions) Axiovert 40c, by Zeiss (1 mentions) Pv 820 pico injector, by World Precision Instruments (1 mentions)

Close spelling variants of this product

MMESSAGE mMACHINE Kit High Yield Capped RNA Transcription Kit MMESSAGE mMACHINE High Yield Capped RNA Transcription kit MMESSAGE mMACHINE high yield capped RNA kit MMessage high-yield capped RNA transcription kit MMESSAGE mMACHINE capped RNA transcription kit SP6 mMessage mMachine MMessage mMachine kit MMACHINE SP6 Kit SP6 mMessage kit MMessage mMachine

2 protocols using sp6 mmessage mmachine high yield capped rna transcription kit

Platynereis Zygote Microinjection Protocol

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Platynereis zygotes were injected using a Zeiss Axiovert 40C inverted microscope, equipped with a joystick micromanipulator (Narishige) and a microinjector (FemtoJet, Eppendorf). Injection needles were pooled from glass capillaries (borosilicate thin wall with filament, 1.0 mm outer diameter, 0.78 mm inner diameter, Harvard apparatus) using a Sutter needle pooler. Embryos were accommodated on an agarose stage, customized for Platynereis injections.
GCaMP6s (Chen et al., 2013 (link)) was kindly provided by the GENIE Project, Janelia Farm Research Campus, Howard Hughes Medical Institute (Addgene plasmid 40753). The GCaMP6s coding sequence was subcloned in the pCS2+ vector, and mRNA was in vitro transcribed with the SP6 mMessage mMachine High Yield Capped RNA Transcription Kit (Ambion). GCaMP6s mRNA was injected in the CD blastomere to a final concentration of 250 μg/μl. H2B-RFP mRNA was co-injected at the same concentration in order to label the CD lineage.

Tosches M.A., Bucher D., Vopalensky P, & Arendt D. (2014). Melatonin Signaling Controls Circadian Swimming Behavior in Marine Zooplankton. Cell, 159(1), 46-57.

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Spatiotemporal Regulation of Smoc1 in Zebrafish Embryos

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One-cell stage wildtype AB or Smoc1^ug104; BRE:GFP; H2B-mCherry embryos were injected using standard procedures with different smoc1 mRNA concentrations. This was generated by linearization of pCS2-smoc1 vector with NotI (NEB), and transcribed using the SP6 mMESSAGE mMACHINE High Yield Capped RNA Transcription Kit (Ambion). The same procedure was performed to obtain smoc1-mCherry, smoc1^ug104-mCherry and Cas9 mRNAs. For antisense morpholino oligonucleotide (MO) injections, 2ng or 12ng of a splicing block MO (5′-CCGGAACTCTGACAGACCTGAGCAA-3′) (Abouzeid et al., 2011 (link)) and its respective mismatch control MO (5′-CCGCAAGTCTCACACACCTCAGCAA-3′) were used (Gene Tools, LLC) in BRE:GFP; H2B-mCherry heterozygote embryos. Embryos were left to develop at 28.5°C until the desired stage and then processed for SEM, live imaging or immunostainings. A PV-820 Pico-injector (World Precision Instruments) and a Narashige micromanipulator were used for microinjection.

Mateus R., Holtzer L., Seum C., Hadjivasiliou Z., Dubois M., Jülicher F, & Gonzalez-Gaitan M. (2020). BMP Signaling Gradient Scaling in the Zebrafish Pectoral Fin. Cell Reports, 30(12), 4292-4302.e7.

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