Superscript rt kit

Manufactured by Thermo Fisher Scientific

Sourced in United States, Canada

The SuperScript RT kit is a laboratory product for reverse transcription, which is the process of converting RNA into complementary DNA (cDNA). The kit contains enzymes, buffers, and other reagents required for this reaction.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

RT kits (5 citations)

38 protocols using superscript rt kit

Quantitative Analysis of SR-B1 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNA from NB cell lines SK-N-Be(2), IMR32, SH-EP, SH-SY5Y, SK-N-As, the positive control NCI-H295R, and the negative control (Jurkat and fibroblast cells) were extracted using Qiagen RNA isolation kit (Qiagen Sciences, Valencia, CA). 500 ng of RNA was reverse transcribed using superscript RT kit from Life Technologies. Quantitative polymerase chain reaction (PCR) was performed in a step-1 real-time PCR (RT-PCR) machine using SR-B1 and actin specific primer sets. Relative gene expression levels were calculated after normalization with internal controls. The expression of SR-B1 was further confirmed at the protein level by western blot analysis as previously described and actin was used as a loading control (12 (link)).

Subramanian C., White P., Kuai R., Avinaash K., Castle V., Moon J., Timmermann B., Schwendeman A, & Cohen M. (2018). Synthetic HDL Nanoconjugate Targets Neuroblastoma Stem Cells, Blocking Migration and Self-Renewal. Surgery.

+ Open protocol

+ Expand

Quantifying TF Gene Expression in HUVECs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted from HUVECs using the RNeasy mini kit (Qiagen). The total RNA (1 μg) was reverse‐transcribed into cDNA using the SuperScript RT kit (Life Technologies), and real‐time polymerase chain reaction analysis was performed on a StepOne system (Applied Biosystems) using TaqMan probes and primers for human TF (assay ID; Hs01076032_m1). Human hypoxanthine phosphoribosyltransferase 1 (assay ID; Hs02800695_m1) served as an endogenous control.

Morimoto M., Tatsumi K., Yuui K., Terazawa I., Kudo R, & Kasuda S. (2021). Convallatoxin, the primary cardiac glycoside in lily of the valley (Convallaria majalis), induces tissue factor expression in endothelial cells. Veterinary Medicine and Science, 7(6), 2440-2444.

+ Open protocol

+ Expand

Transcriptional Regulation Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Myc, YAP, and Scribbled expression was analyzed using the Superscript RT kit (Life Technologies 18080–051) and the SSoadvanced SYBRGreen qPCR mix (BioRad 172–5260) with the following primers pairs: MYC (GTAGTGGAAAACCAGCAGCCT, AGAAATACGGCTGCACCGAG); YAP (TGACCCTCGTTTTGCCATGA, GTTGCTGCTGGTTGGAGTTG) Scribbled (AGGAGATCTACCGCTACAG, GATCTCAGGGATATCGTTCC); HIF1A (GTGAAGACATCGCGGGGA, GTGGCAACTGATGAGCAAGC); GAPDH (GGTGAAGGTCGGAGTCAACGG, GAGGTCAATGAAGGGGTCATTG).

Penzo-Méndez A.I., Chen Y.J., Li J., Witze E.S, & Stanger B.Z. (2015). Spontaneous Cell Competition in Immortalized Mammalian Cell Lines. PLoS ONE, 10(7), e0132437.

+ Open protocol

+ Expand

Quantification of SR-BI Expression in Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNA from the NCI-H295R cells after drug treatment for 24 h was prepared using Qiagen RNA isolation kit (Qiagen Sciences, Valencia, CA). Approximately 500 ng of RNA was reverse transcribed using superscript RT kit from Life Technologies (Grand Island, NY). qPCR was performed in a step-one RT PCR machine using the gene specific primer sets (Table 1A) as published (18 (link)). Relative gene expression levels were calculated after normalization with internal controls. SR-BI expression level in several cancers was confirmed by Western-Blot Analysis(Table 1B).

Subramanian C., Kuai R., Zhu Q., White P., Moon J., Schwendeman A, & Cohen M.S. (2015). sHDL Nanoparticles: A Novel Therapeutic Strategy for Adrenocortical Carcinomas. Surgery, 159(1), 284-295.

+ Open protocol

+ Expand

RNA Extraction and Real-Time PCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was isolated from cells with TRIzol reagent (Invitrogen). The SuperScript RT kit (Fermentas, Ottawa, Canada) was adopted to synthesize cDNA. The RT-qPCR assay was then performed on the Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Foster City, USA) by utilizing SYBR Green PCR Master Mix (Takara Bio, Otsu, Japan). U6 and GAPDH were used as internal references, respectively. RNA expression was assessed using 2^-ΔΔCt method. The primers were shown in Table 1.

Wang W., Yi J., Dong D., Mao W., Wang X, & Yan Z. (2021). miRNA-877-5p inhibits malignant progression of prostate cancer by directly targeting SSFA2. European Journal of Histochemistry : EJH, 65(3), 3243.

+ Open protocol

+ Expand

RT-qPCR Analysis of miR-4269, ZEB1, and OTX1

Check if the same lab product or an alternative is used in the 5 most similar protocols

According to the supplier’s instructions, total RNA was isolated from PC cells with TRIzol reagent (Invitrogen). The SuperScript RT kit (Fermentas, Ottawa, Canada) was adopted to synthesize cDNA. The RT-qPCR assay was then performed on a Applied Biosystems 7500 Real-Time PCR system (Applied Biosystems, Foster City, U.S.A.) by utilizing SYBR Green PCR Master Mix (Takara Bio, Otsu, Japan). The 2^−ΔΔCT method was adopted to calculate gene expression levels. GAPDH and U6 were employed as internal controls for normalization. Primers were shown as follows: miR-4269: 5′-GCAGGCACAGACAGCCCTG-3′ (sense) and 5′-GAACATGTCTGCGTATCTC-3′ (antisense); ZEB1: 5′-GATGACCTGCCAACAGACCA′ (sense) and 5′-CCCCAGGATTTCTTGCCCTT-3′ (antisense); OTX1: 5′-CTGCTCTTCCTCAATCAATGG-3′ (sense) and 5′-ACCCTGACTTGTCTGTTTCC-3′ (antisense); U6: 5′-GCTTCGGCAGCACATATACTAAAAT-3′ (sense) and 5′-CGCTTCAGAATTTGCGTGTCAT-3′ (antisense).

Sui X, & Sui Z. (2020). MiR-4269 suppresses the tumorigenesis and development of pancreatic cancer by targeting ZEB1/OTX1 pathway. Bioscience Reports, 40(6), BSR20200010.

+ Open protocol

+ Expand

Quantitative Analysis of Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

TRIzol reagent (Invitrogen, USA) was used to collect total RNA from cells and tissues. The RNA was reversely transcribed into cDNA using a SuperScriptRT Kit (Invitrogen, USA). Then, the cDNA was collected for real-time qPCR using the SYBR Green Master Mix (Applied Biosystems Inc., Carlsbad, CA, USA) on an ABI PRISM 7300 RT-PCR System (Applied Biosystems). The primer sequences are listed in Table 1, and the relative RNA expression was determined using the 2^−ΔΔCt method with U6 and GAPDH as the internal references for miRNA and mRNAs, respectively.Table 1

Primer Sequences in RT-qPCR

Gene	Primer Sequence (5ʹ-3ʹ)
miR-125a	F: TCCCTGAGACCCTTTAACCT
miR-125a	R: GAACATGTCTGCGTATCTC
HDAC1	F: TGAAGCCTCACCGAATCCGCAT
HDAC1	R: TGGTCATCTCCTCAGCATTGGC
ET-1	F: CTACTTCTGCCACCTGGACATC
ET-1	R: CGCACTGACATCTAACTGCCTG
Col. I	F: CCTCAGGGTATTGCTGGACAAC
Col. I	R: CAGAAGGACCTTGTTTGCCAGG
FN	F: CCCTATCTCTGATACCGTTGTCC
FN	R: TGCCGCAACTACTGTGATTCGG
IL-6	F: TACCACTTCACAAGTCGGAGGC
IL-6	R: CTGCAAGTGCATCATCGTTGTTC
GAPDH	F: GCACCGTCAAGGCTGAGAAC
GAPDH	R: TGGTGAAGACGCCAGTGGA
U6	F: GCTTCGGCAGCACATATACTAAAA
U6	R: GCTTCGGCAGCACATATACTAAAAT

Abbreviations: RT-qPCR, reverse transcription quantitative polymerase chain reaction; HDAC1, histone deacetylase 1; ET-1, endothelin-1; Col. I, collagen I; FN, fibronectin; IL-6, interleukin-6; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Hao Y., Miao J., Liu W., Cai K., Huang X, & Peng L. (2021). Mesenchymal Stem Cell-Derived Exosomes Carry MicroRNA-125a to Protect Against Diabetic Nephropathy by Targeting Histone Deacetylase 1 and Downregulating Endothelin-1. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 14, 1405-1418.

+ Open protocol

+ Expand

Quantitative Real-Time PCR Analysis of ZNRF3

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA was extracted from NPC cell lines using TRIzol reagent (Takara, Dalian, P.R. China) following the manufacturer’s instructions, and 5 μg of RNA of each sample was reverse transcribed using SuperScript RT kit (Invitrogen). All the qPCRs were performed on a StepOne™ Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The following primers were used: ZNRF3, 5′-GCGGGTCATCCCCTGTAC-3′ (sense) and 5′-GCTTGGGTTTCCCTTTTGTT-3′ (antisense); β-actin, 5′-CTTAGTTGCGTTACACCCTTTCTTG-3′ (sense) and 5′-CTGTCACCTTCACCGTTCCAGTTT-3′ (antisense). The band intensities of amplification products were measured by a densitometer, and the results were normalized with β-actin.

Wang Z., Wang Y., Ren H., Jin Y, & Guo Y. (2017). ZNRF3 Inhibits the Invasion and Tumorigenesis in Nasopharyngeal Carcinoma Cells by Inactivating the Wnt/β-Catenin Pathway. Oncology Research, 25(4), 571-577.

+ Open protocol

+ Expand

Total RNA Extraction and qRT-PCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The extraction of total RNA from cells was performed by Trizol reagent (Life Technologies Corporation, Carlsbad, CA, USA) following directions. The Taqman MicroRNA Reverse Transcription Kit and Taqman Universal Master Mix II with the TaqMan MicroRNA Assay of miR-203 and U6 (Applied Biosystems, Foster City, CA, USA) were used to detect expression level of miR-203 in cells. In addition, for inflammatory factors (TNF-α and IL-6) and inducible NOS (iNOS), the SuperScript RT kit (Invitrogen, Carlsbad, CA, USA) was used to reverse transcription of RNA, and the 7500c real-time PCR detection system (Applied Biosystems, Carlsbad, CA, USA) with SYBR premix EX Taq (TaKaRa) was used to detect expression levels of mRNA, with β-actin as internal control.

Li D., Li G., Chen Y., Li Y., Zhang J., Gao D., Sun L, & Liu B. (2020). Astragaloside IV protects ATDC5 cells from lipopolysaccharide-caused damage through regulating miR-203/MyD88. Pharmaceutical Biology, 58(1), 89-97.

+ Open protocol

+ Expand

qRT-PCR Analysis of Gene Expression

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

qRT-PCR was performed as described (Spannl et al., 2017 (link)). Briefly, RNA extraction was performed using a standard Trizol-based (Invitrogen) procedure with phase separation using chloroform according to the manufacturer’s instructions. Approximately 10 heads or 40–50 dissected eyes per genotype constituted one biological replicate, and at least two to five biological replicates were used. cDNA generation was performed using SuperScript RT kit (Invitrogen) with a starting amount of 1 µg total RNA. qPCR was performed using primers described in Table S4.

Hebbar S., Schuhmann K., Shevchenko A, & Knust E. (2020). Hydroxylated sphingolipid biosynthesis regulates photoreceptor apical domain morphogenesis. The Journal of Cell Biology, 219(12), e201911100.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!