RNA extraction and real-time PCR was conducted as described previously (Morrison et al., 2007 (link)). Total RNA was extracted from liver, iWAT, and BAT using TRIzol reagent following the manufacturer’s protocol (15596018, Invitrogen). RNA quality and quantity was determined by spectrophotometry using a NanoDrop (Thermo Scientific). cDNA synthesis was performed with M-MLV reverse transcriptase (M1701, Promega) and mRNA was quantified on the ABI 7900 platform using the SYBR green methodology in optical 384-well plates (Applied Biosystems). Primer pairs were designed using NCBI Primer-BLAST with at least one primer spanning an exon-exon boundary. Target gene expression was normalized with cylcophilin B as the endogenous control.
Abi 7900 platform
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 7900 platform is a real-time PCR system designed for gene expression analysis. It is capable of detecting and quantifying nucleic acid sequences.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop, by Thermo Fisher Scientific (4 mentions)
Pcr master mix, by Thermo Fisher Scientific (2 mentions)
Taqman gene expression cells to ct kit, by Thermo Fisher Scientific (2 mentions)
Optical 384 well plates, by Thermo Fisher Scientific (1 mentions)
M mlv reverse transcriptase, by Promega (1 mentions)
Abi sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Iscript, by Bio-Rad (1 mentions)
Rneasy lipid tissue mini kit, by Qiagen (1 mentions)
Applied biosystems high capacity reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Sybr premix ex taq, by Takara Bio (1 mentions)
Primescript rt reagent kit with gdna eraser, by Takara Bio (1 mentions)
Sybr premix ex taq 2, by Takara Bio (1 mentions)
Rnaiso plus kit, by Takara Bio (1 mentions)
7 protocols using abi 7900 platform
1
RNA Extraction and Real-Time PCR Protocol
2
RNA Extraction and Real-Time PCR for Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA extraction and real-time PCR were conducted as described previously36 (link),42 (link). Total RNA was extracted from liver and inguinal white adipose tissue (iWAT) using TRIzol reagent following the manufacturer’s protocol (Invitrogen), with the addition of an RNeasy Lipid Tissue Mini Kit (QIAGEN) for iWAT. RNA purity and quantity were determined by spectrophotometry using a NanoDrop (Thermo Scientific). cDNA synthesis was performed with iScript (BioRad) and mRNA was quantified on the ABI 7900 platform using the ABI SYBR Green PCR Master Mix in optical 384-well plates (Applied Biosystems). Primer pairs were designed using the IDT RealTime qPCR Primer Design tool to span an intro-exon boundary (Table S3 ). Target gene expression was normalized with cyclophilin as the endogenous control.
3
MKL-1 Cell RNA Isolation and qPCR
4
Gene Expression Analysis in Tumor Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We isolated RNA from cells and crushed tumor tissue samples using the TRIZOL method. cDNA was obtained using the PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa, Cat# RR047A) according to the manufacturer’s instructions. SYBR Premix Ex Taq (TaKaRa, Cat# RR420A) was used to analyze gene expression by RT‐PCR carried out using the ABI 7900 platform (Applied Biosystems). Relative mRNA levels were normalized to β‐actin mRNA levels. The primers used are included in Table S2 .
5
HKII Expression Analysis in Resistant Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HKII gene expression level in the parental and resistant cells was analyzed by quantitative real-time polymerase chain reaction (qPCR). Total RNA was extracted and converted in complementary Deoxyribonucleic acid (DNA) using the Taqman Gene Expression cells-to-CT kit (Life Technologies, Grand Island, NY). Real-time PCR reaction was carried out using PCR Master Mix (Life Technologies) on ABI-7900 platform (Applied Bio systems). HKII was amplified from cDNA using primers designed to amplify its coding region (Applied Bio systems, Hs00606086).
6
Gene Expression Profiling in Lymphoma
7
Quantifying NEAT1 Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted using the RNAiso Plus kit (Takara). Reverse transcription of RNA was performed using TM RT Master Mix (Takara), and qPCR was performed using SYBR Premix Ex Taq II (Takara). The qRT-PCR reaction was performed on an ABI 7900 platform (Applied Biosystems). mRNA expression was determined with β-actin as an internal reference and the relative expression level of the target gene was calculated using the 2 -△△CT method. Primer sequences are as follows: NEAT1-F: TTG TTC CAG AGC CCA TGA T; NEAT1-R: TGA AAA CCT TTA CCC CAG GA; β-actin-F: GGC TCC GGC ATG TGC AAG; β-actin-R: CCT CGG TCA GCA GCA CGG .
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!