Iscript reverse transcription supermix for qrt pcr
The IScript Reverse Transcription Supermix for qRT-PCR is a reagent used for the reverse transcription of RNA to cDNA, which can then be used for quantitative real-time PCR (qRT-PCR) analysis. It contains all the necessary components, including reverse transcriptase, RNase inhibitor, and optimized buffer, in a single ready-to-use solution.
Lab products found in correlation
33 protocols using iscript reverse transcription supermix for qrt pcr
Quantifying RNA Expression Using Direct-zol Extraction
Gene Expression Analysis of Immune Markers
Sequences of PCR primers used in this study
Gene | Forward | Reverse |
---|---|---|
CD147 | 5′-GACACTGGGGAAGAAGAGGC-3′ | 5′-GCAGTGAGATGGTTTCCCGA-3′ |
TNF-α | 5′-AGTCCGGGCAGGTCTACTTT-3′ | 5′-ACCCTGAGCCATAATCCCCT-3′ |
IL-1β | 5′-ATGCCACCTTTTGACAGTGATG-3′ | 5′-GCAGCCCTTCATCTTTTGGG-3′ |
IL-6 | 5′-CCTTCCAGGATGAGGACATGA-3′ | 5′-TGAGTCACAGAGGATGGGCTC-3′ |
Arg1 | 5′-TCACCTGAGCTTTGATGTCG-3′ | 5′- CTGAAAGGAGCCCTGTCTTG-3′ |
MCP-1 | 5′-CCCCAAGAAGGAATGGGTCC-3′ | 5′-GTGCTGAAGACCTTAGGGCA-3′ |
Ly6C | 5′-ACTGTGCCTGCAACCTTGT-3′ | 5′-GCTGGGCAGGAAGTCTCAAT-3′ |
CCR2 | 5′-ATCCACGGCATACTATCAACATC-3′ | 5′-CAAGGCTCACCATCATCGTAG-3′ |
iNOS | 5′-CAAGCACCTTGGAAGAGGAG-3′ | 5′- AAGGCCAAACACAGCATACC-3′ |
GAPDH | 5′-GCGAGATCCCGCTAACATCA-3′ | 5′-CTCGTGGTTCACACCCATCA-3′ |
RNA Isolation and qRT-PCR Analysis
RNA Extraction and cDNA Synthesis from Liver and Skeletal Muscle Tissues
Quantitative RT-PCR Assay for CHEK1 and RAD54L
RNA Extraction and qRT-PCR Analysis
Quantitative Analysis of CDK2 Expression
HPV16 E6 Gene Expression Analysis
Quantitative RT-PCR Analysis of Gene Expression
Quantitative RT-PCR Analysis of HER2 in Metastatic Breast Cancer
Example 19
Total RNA was extracted from the 21MT-1 metastatic breast cancer cell line at 72 hours post transfection using Trizol (Life Technologies) and following standard RNA extraction protocol. RNA quantity and quality was assessed by a NanoDrop (Thermo Scientific) UV/VIS spectrophotometer, cDNA was made with iScript Reverse Transcription Supermix for qRT-PCR (Bio-Rad), and a RealPlex thermocycler (Eppendorf) was used to create cDNA (5 min 25° C., 30 min 42° C., 5 min 85° C.). Following cDNA creation, qRT-PCR was performed by using the SYBR Green Master Mix (Life Technologies), and a per-reaction use of 500 nM of both the forward and reverse primers with 10 ng of cDNA. The qRT-PCR was performed with a 40 cycle amplification (95° C. for 10 seconds, 56° C. for 30 seconds, and 72° C. for 30 seconds) and a threshold intensity of 50 (during linear phase). The primers used for the qRT-PCR analysis are as follows:
The ΔΔCt Method was used to calculate changes in gene expression, and at least three biological samples were analyzed.
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