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Iscript reverse transcriptase supermix

Manufactured by Thermo Fisher Scientific
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The IScript Reverse Transcriptase Supermix is a ready-to-use solution for reverse transcription of RNA into cDNA. It contains all the necessary components for efficient and reliable cDNA synthesis.

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Lab products found in correlation

Nucleospin rna extraction kit, by Takara Bio (2 mentions) Fast sybr green mix, by Thermo Fisher Scientific (2 mentions) Ab9110, by Abcam (1 mentions) Image lab software, by Bio-Rad (1 mentions)

2 protocols using iscript reverse transcriptase supermix

1

Investigating Gmnn-Zic1 Interaction in HEK 293 Cells

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Total RNA was extracted using Clontech Nucleospin RNA extraction kit (#740955.250) using the manufacturer’s protocol. cDNA was synthesized using the Bio Rad iScript Reverse Transcriptase Supermix (#170-8841) followed by quantitative RT-PCR analysis using the Fast SYBR green mix from Applied Biosystems (#4385612) and gene-specific primer sequences shown in Supplementary Table S5. Anti-Gmnn antibody from Santa Cruz (Cat. No. #sc-13015) and anti-Zic1 antibody from Rockland Inc. (Cat. #200-401-159) were used for immunoblotting at 1:200 dilution. The Gmnn and Zic1 bands were quantified using Bio-Rad Image Lab software. Co-immunoprecipitation experiments were performed as previously described16 (link) in HEK 293 cells transfected with HA-tagged Gmnn and Myc-tagged Zic1. Immunoprecipitation was performed with the HA tag using HA antibody (Abcam: ab9110) and blotted for Myc using Myc antibody (Abcam: ab32).
Sankar S., Yellajoshyula D., Zhang B., Teets B., Rockweiler N, & Kroll K.L. (2016). Gene regulatory networks in neural cell fate acquisition from genome-wide chromatin association of Geminin and Zic1. Scientific Reports, 6, 37412.
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2

RNA Extraction, cDNA Synthesis, and qRT-PCR Analysis

Cited 1 time
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Total RNA was extracted with the Clontech Nucleospin RNA extraction kit (#740955.250), using the manufacturer's protocol. cDNA was synthesized using the Bio Rad iScript Reverse Transcriptase Supermix (#170-8841), followed by quantitative RT-PCR analysis using the Fast SYBR green mix from Applied Biosystems (#4385612), as previously described [34 (link)]. Primers used were: mouse Gmnn (F-GCAGTACATGGCGGAGGTAATC and R-GCTCCTGAGTCTTCCAGTTCTG), mouse α-Actin (F-CATTGCTGACAGGATGCAGAAGG and R-TGCTGGAAGGTGGACAGTGAGG), human Gmnn (F-CAGCCTTCTGCATCTGGATCTC and R-CCAGGGCTGGAAGTTGTAGATG), human GAPDH (F-GTCTCCTCTGACTTCAACAGCG and R-ACCACCCTGTTGCTGTAGCCAA), human α-Actin (F-CACCATTGGCAATGAGCGGTTC and R-AGGTCTTTGCGGATGTCCACGT). Protein lysate preparation and Western blotting was performed as previously described [30 (link)].
Sankar S., Patterson E., Lewis E.M., Waller L.E., Tong C., Dearborn J., Wozniak D., Rubin J.B, & Kroll K.L. (2017). Geminin deficiency enhances survival in a murine medulloblastoma model by inducing apoptosis of preneoplastic granule neuron precursors. Genes & Cancer, 8(9-10), 725-744.
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