Sp6 t7 t3 polymerases

Manufactured by Roche

SP6/T7/T3 polymerases are enzymes used for in vitro transcription. They catalyze the synthesis of RNA from DNA templates containing the respective promoter sequences.

Automatically generated - may contain errors

Lab products found in correlation

Dig labelling mix, by Roche (2 mentions) Noti restriction enzyme, by New England Biolabs (2 mentions) Sacii, by New England Biolabs (2 mentions)

Close spelling variants of this product

SP6 polymerase T3 polymerase T7 polymerase

2 protocols using sp6 t7 t3 polymerases

Synthetic mRNA and sgRNA Production for Cell Labeling

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Synthetic capped mRNAs were transcribed using the SP6 mMessage mMachine, following the manufacturer’s instructions. Cas9 sgRNA was transcribed using the T7 HiScribe High Yield RNA Synthesis kit, following the manufacturer’s instructions. RNAs were purified with phenol-chloroform extraction. The pSP64-mmGFP5-nos1-3’UTR [48 (link)] and pSP64-eGFP-F-nos1-3’UTR [49 (link)] constructs were linearised with SacII and NotI restriction enzymes (NEB) respectively, following the manufacturer’s instructions. These were used to label the cytoplasm and cell membranes of the PGCs. Probes for whole mount in situ hybridisation were produced with Promega SP6/T7/T3 polymerases with DIG-labelling mix (Roche) and purified with lithium chloride extraction.

Vong Y.H., Sivashanmugam L., Leech R., Zaucker A., Jones A, & Sampath K. (2021). The RNA-binding protein Igf2bp3 is critical for embryonic and germline development in zebrafish. PLoS Genetics, 17(7), e1009667.

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mRNA Synthesis and Visualization Protocols

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Synthetic capped mRNAs were transcribed using the SP6 mMessage mMachine, following the manufacturer's instructions. Cas9 sgRNA was transcribed using the T7 HiScribe High Yield RNA Synthesis kit, following the manufacturer's instructions. RNAs were purified with phenol-chloroform extraction. The pSP64-mmGFP5-nos1-3'UTR (Köprunner et al., 2001 (link)) and pSP64-eGFP-F-nos1-3'UTR (Weidinger et al., 2002) constructs were linearised with SacII and NotI restriction enzymes (NEB) respectively, following the manufacturer's instructions. These were used to label the cytoplasm and cell membranes of the PGCs.
Probes for whole mount in situ hybridisation were produced with Promega SP6/T7/T3 polymerases with DIG-labelling mix (Roche) and purified with lithium chloride extraction.

Vong Y.H., Sivashanmugam L., Zaucker A., Jones A.M.E., & Sampath K. (2020). The RNA-binding protein Igf2bp3 is critical for embryonic and germline development in zebrafish.

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