MPRA template was in vitro transcribed, capped, tailed, and purified using the mMESSAGE mMACHINE T7 ULTRA Transcription Kit (Invitrogen). Eleven reactions in total were carried out, each using 1μg of MPRA template in the manufacturer recommended reaction with the addition of 1μLSUPERase-In RNase Inhibitor (Invitrogen). Reactions were incubated at 37°C for 4 h, then 1μL Turbo DNase (Invitrogen) added and incubated at 37°C for an additional 15 min for removal of DNA template. Poly-A tailing protocol was followed to manufacturer’s instructions with a 45-min incubation at 37°C. RNA recovery was performed using lithium chloride precipitation according to kit protocol. All eleven reaction products (~350μg) were pooled to make the MPRA mRNA library.
μlsuperase in rnase inhibitor
Manufactured by Thermo Fisher Scientific
μLSUPERase-In RNase Inhibitor is a recombinant RNase inhibitor protein that protects RNA from degradation by RNase enzymes during experimental procedures. It provides highly effective and stable RNase inhibition, ensuring the integrity of RNA samples.
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Lab products found in correlation
2 protocols using μlsuperase in rnase inhibitor
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High-Yield MPRA mRNA Library Synthesis
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High-Yield MPRA mRNA Library Synthesis
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MPRA template was in vitro transcribed, capped, tailed, and purified using the mMESSAGE mMACHINE T7 ULTRA Transcription Kit (Invitrogen). Eleven reactions in total were carried out, each using 1μg of MPRA template in the manufacturer recommended reaction with the addition of 1μLSUPERase-In RNase Inhibitor (Invitrogen). Reactions were incubated at 37°C for 4 h, then 1μL Turbo DNase (Invitrogen) added and incubated at 37°C for an additional 15 min for removal of DNA template. Poly-A tailing protocol was followed to manufacturer’s instructions with a 45-min incubation at 37°C. RNA recovery was performed using lithium chloride precipitation according to kit protocol. All eleven reaction products (~350μg) were pooled to make the MPRA mRNA library.
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