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Superscript 3 first strand synthesis system for qrt pcr protocol

Manufactured by Thermo Fisher Scientific
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The SuperScript III First-Strand Synthesis System for qRT-PCR protocol is a reverse transcription kit designed for the synthesis of first-strand cDNA from total RNA or poly(A)+ RNA templates. The system utilizes the SuperScript III reverse transcriptase enzyme, which provides improved sensitivity and reduced RNase H activity. The kit includes components for first-strand cDNA synthesis, along with optional random hexamers or oligo(dT) primers for priming the reaction.

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2 protocols using superscript 3 first strand synthesis system for qrt pcr protocol

1

Validating MSNP1AS Knockdown via qRT-PCR

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To confirm knockdown of MSNP1AS, cDNA was synthesized using the SuperScript III First-Strand Synthesis System for qRT-PCR protocol (Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA). Five hundred nanograms of RNA were used to make one and a half reaction volumes (35 µL) of cDNA. The cDNA was stored at −20 °C. The qRT-PCR protocol described in Kerin et al. [7 ] was used to validate knockdown of MSNP1AS.
DeWitt J.J., Hecht P.M., Grepo N., Wilkinson B., Evgrafov O.V., Morris K.V., Knowles J.A, & Campbell D.B. (2016). Transcriptional Gene Silencing of the Autism-Associated Long Noncoding RNA MSNP1AS in Human Neural Progenitor Cells. Developmental neuroscience, 38(5), 375-383.
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2

Validating MSNP1AS Overexpression in ASD

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To confirm overexpression of MSNP1AS, consistent with the ~12-fold increase observed in postmortem brains of individuals with ASD, cDNA was synthesized using the SuperScript III First-Strand Synthesis System for qRT-PCR protocol (Invitrogen/Thermo Fisher Scientific, Waltham, MA, USA). Five hundred nanograms of RNA were used to make one and a half reaction volumes (35 μL) of cDNA. The cDNA was stored at −20 °C. The qRT-PCR protocol described in Kerin et al. [9 (link)] was used to validate over-expression of MSNP1AS.
DeWitt J.J., Grepo N., Wilkinson B., Evgrafov O.V., Knowles J.A, & Campbell D.B. (2016). Impact of the Autism-Associated Long Noncoding RNA MSNP1AS on Neuronal Architecture and Gene Expression in Human Neural Progenitor Cells. Genes, 7(10), 76.
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