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Superscript 3 strand synthesis system

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The Superscript III strand synthesis system is a laboratory instrument designed for the synthesis of complementary DNA (cDNA) from RNA templates. The system utilizes reverse transcriptase enzymes to facilitate the conversion of RNA to single-stranded cDNA, which can then be used in various downstream applications such as gene expression analysis, cloning, and library construction.

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Lab products found in correlation

Mastercycler, by Eppendorf (3 mentions) Sybr green based detection, by Thermo Fisher Scientific (3 mentions) Trizol, by Thermo Fisher Scientific (3 mentions) Steponeplus platform, by Thermo Fisher Scientific (1 mentions) Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Superscript III (4906 citations) SuperScript III First-Strand Synthesis System kit (143 citations) SuperScript III system (137 citations) SuperScript III First-Strand Synthesis System for RT-PCR kit (71 citations) SuperScript III First-Strand Synthesis System for reverse transcription-PCR (19 citations) SuperScript III synthesis system (14 citations) Superscript III First Strand Synthesis System for RT-PCR kit with oligo (dT) primers (4 citations) SuperScript III first-strand synthesis system for real-time PCR (4 citations) SuperScript III Synthesis (2 citations) Invitrogen SuperScript® III First-Strand Synthesis System for RT-PCR (2 citations)

4 protocols using superscript 3 strand synthesis system

1

Quantitative Analysis of POLA1 Expression

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RNA was extracted using Trizol (Invitrogen) according to the manufacturer’s instructions. RNA (5μg) was used for cDNA synthesis utilizing the Superscript III strand synthesis system (Invitrogen). For qualitative analysis of POLA1 transcripts, the following primer sequences were utilized: primer A (exon 10, 5’-AAAGGGGCAGATGAGGAACAA-3’); primer X (exon 13a, 5’-TCTGACAGTGGTGATGAAAAG-3’); primer B (exon 15, 5’-ACAAG CGGTGGTGGACTGAC-3’). Quantitative real-time RT-PCR was performed using SYBR Green based detection (Invitrogen) and a Mastercycler (Eppendorf, Germany) as previously reported46 (link). Experiments were performed using technical duplicates or triplicates, data were normalized to housekeeping genes and the relative abundance of transcripts was calculated by the comparative ΔΔCt method. All primers used for qRT-PCR are indicated in Supplementary Table 6.
Starokadomskyy P., Gemelli T., Rios J.J., Xing C., Wang R.C., Li H., Pokatayev V., Dozmorov I., Khan S., Miyata N., Fraile G., Raj P., Xu Z., Xu Z., Ma L., Lin Z., Wang H., Yang Y., Ben-Amitai D., Orenstein N., Mussaffi H., Baselga E., Tadini G., Grunebaum E., Sarajlija A., Krzewski K., Wakeland E.K., Yan N., de la Morena M.T., Zinn A.R, & Burstein E. (2016). DNA polymerase-α regulates type I interferon activation through cytosolic RNA:DNA synthesis. Nature immunology, 17(5), 495-504.
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2

Quantitative Analysis of POLA1 Expression

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RNA was extracted using Trizol (Invitrogen) according to the manufacturer’s instructions. RNA (5μg) was used for cDNA synthesis utilizing the Superscript III strand synthesis system (Invitrogen). For qualitative analysis of POLA1 transcripts, the following primer sequences were utilized: primer A (exon 10, 5’-AAAGGGGCAGATGAGGAACAA-3’); primer X (exon 13a, 5’-TCTGACAGTGGTGATGAAAAG-3’); primer B (exon 15, 5’-ACAAG CGGTGGTGGACTGAC-3’). Quantitative real-time RT-PCR was performed using SYBR Green based detection (Invitrogen) and a Mastercycler (Eppendorf, Germany) as previously reported46 (link). Experiments were performed using technical duplicates or triplicates, data were normalized to housekeeping genes and the relative abundance of transcripts was calculated by the comparative ΔΔCt method. All primers used for qRT-PCR are indicated in Supplementary Table 6.
Starokadomskyy P., Gemelli T., Rios J.J., Xing C., Wang R.C., Li H., Pokatayev V., Dozmorov I., Khan S., Miyata N., Fraile G., Raj P., Xu Z., Xu Z., Ma L., Lin Z., Wang H., Yang Y., Ben-Amitai D., Orenstein N., Mussaffi H., Baselga E., Tadini G., Grunebaum E., Sarajlija A., Krzewski K., Wakeland E.K., Yan N., de la Morena M.T., Zinn A.R, & Burstein E. (2016). DNA polymerase-α regulates type I interferon activation through cytosolic RNA:DNA synthesis. Nature immunology, 17(5), 495-504.
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3

Quantitative RT-PCR Analysis of Gene Expression

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RNA was extracted using TRIzol (Invitrogen, Thermo Fisher Scientific) according to the manufacturer’s instructions. Total RNA (3 μg) was used for cDNA synthesis utilizing the Superscript III strand synthesis system (Invitrogen, Thermo Fisher Scientific). qRT-PCR was performed using a Mastercycler (Eppendorf) following the manufacturer’s recommendations. SYBR Green–based detection (Invitrogen, Thermo Fisher Scientific) was employed using gene-specific primers noted in Table 4. Experiments were performed in duplicate, data were normalized to housekeeping genes (ACTB, GAPDH, 18S rRNAs, 28S rRNA), and the relative abundance of transcripts was calculated by the comparative ΔΔCt method.
Starokadomskyy P., Wilton K.M., Krzewski K., Lopez A., Sifuentes-Dominguez L., Overlee B., Chen Q., Ray A., Gil-Krzewska A., Peterson M., Kinch L.N., Rohena L., Grunebaum E., Zinn A.R., Grishin N.V., Billadeau D.D, & Burstein E. (2019). NK cell defects in X-linked pigmentary reticulate disorder. JCI Insight, 4(21), e125688.
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4

Quantitative Analysis of Genomic Alterations

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Quantitative polymerase chain reaction (Q-PCR) and a subset of 140 cases for which DNA was available were used to reexamine regions identified as exhibiting aberrant copy number alterations based on the CGH microarray data [28 (link)]. To normalize the copy number per cell, sequences from ß-globin and WNT9A genes were used as endogenous references. Q-PCR for each sample and each gene were performed in triplicate. DNA from three normal tissue samples that were randomly selected from resected specimen was used as the control.
Total RNA was isolated from fresh-frozen tumor samples using the Trizol reagent (Invitrogen, Carlsbad, CS, USA). cDNAs were synthesized from 3 µg of total RNA using the SuperScriptIII-Strand Synthesis System (Invitrogen Japan K.K., Tokyo, Japan). The cDNAs were used for real-time PCR (RT-PCR) analysis for EGFR and ZNF217 expression. RT-PCR reactions were performed on an Applied Biosystems StepOnePlus platform using the TaqMan Fast Advanced Master mix.
Okamoto A., Sehouli J., Yanaihara N., Hirata Y., Braicu I., Kim B.G., Takakura S., Saito M., Yanagida S., Takenaka M., Yamaguchi N., Morikawa A., Tanabe H., Yamada K., Yoshihara K., Enomoto T., Itamochi H., Kigawa J., Matsumura N., Konishi I., Aida S., Aoki Y., Ishii N., Ochiai K., Akiyama T, & Urashima M. (2015). Somatic Copy Number Alterations Associated with Japanese or Endometriosis in Ovarian Clear Cell Adenocarcinoma. PLoS ONE, 10(2), e0116977.
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