Sp6 message mmachine kit

Manufactured by Thermo Fisher Scientific

The SP6 message mMACHINE Kit is a laboratory tool used for the in vitro transcription of messenger RNA (mRNA) from DNA templates. It provides the necessary components for the synthesis of capped and polyadenylated mRNA molecules.

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Lab products found in correlation

Poly a tailing kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

MMACHINE SP6 Kit (9 citations)

3 protocols using sp6 message mmachine kit

mRNA Preparation for Microinjection

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To prepare mRNA for microinjection, the expression vectors were linearized and in vitro transcribed using the SP6 message mMACHINE Kit (Invitrogen, AM1340). Poly(A) tails [~200 to 250 base pairs (bp)] were added to the transcribed mRNAs using the mMACHINE kit (Invitrogen, AM1350). The in vitro–transcribed mRNAs were resuspended in nuclease-free water by lithium chloride precipitation method.

Dai X.X., Pi S.B., Zhao L.W., Wu Y.W., Shen J.L., Zhang S.Y., Sha Q.Q, & Fan H.Y. (2022). PABPN1 functions as a hub in the assembly of nuclear poly(A) domains that are essential for mouse oocyte development. Science Advances, 8(43), eabn9016.

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Preparing mRNA for Microinjection

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To prepare mRNAs for microinjection, expression vectors were linearized, and subjected to phenol/chloroform extraction and ethanol precipitation. Linearized DNAs were in vitro transcribed using the SP6 message mMACHINE Kit (Invitrogen, AM1340). Transcribed mRNAs were added with poly (A) tails (∼200–250 bp) using the mMACHINE Kit (Invitrogen, AM1350), recovered by lithium chloride precipitation, and resuspended in nuclease-free water.

Sha Q.Q., Zhu Y.Z., Li S., Jiang Y., Chen L., Sun X.H., Shen L., Ou X.H, & Fan H.Y. (2019). Characterization of zygotic genome activation-dependent maternal mRNA clearance in mouse. Nucleic Acids Research, 48(2), 879-894.

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In Vitro Transcription and Polyadenylation

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To prepare mRNAs for microinjection, expression vectors were linearized and subjected to phenol/chloroform extraction and ethanol precipitation. The linearized DNAs were transcribed in vitro using the SP6 message mMACHINE Kit (Invitrogen, AM1340). Poly(A) tails (~200–250 bp) were added to transcribed mRNAs using the Poly(A) tailing kit (Invitrogen, AM1350) and were recovered by lithium chloride precipitation and resuspended in nuclease-free water.

Wu Y.W., Li S., Zheng W., Li Y.C., Chen L., Zhou Y., Deng Z.Q., Lin G., Fan H.Y, & Sha Q.Q. (2022). Dynamic mRNA degradome analyses indicate a role of histone H3K4 trimethylation in association with meiosis-coupled mRNA decay in oocyte aging. Nature Communications, 13, 3191.

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