Total RNA was extracted with RNEasy Mini or Micro Plus kits (Qiagen) followed by cDNA synthesis using the Superscript III First Strand Synthesis SuperMix for qRT-PCR kit (Life Technologies). Samples were analyzed on an ABI Fast 7900HT under standard conditions using the TaqMan Gene Expression Assays (Life Technologies). Data were normalized to HRPT1 and analyzed using the comparative CT method.
Fast 7900ht
Manufactured by Thermo Fisher Scientific
The Fast 7900HT is a real-time PCR system designed for high-throughput gene expression analysis. It features a fast thermal cycling capability and supports a range of sample formats, including 384-well and 96-well plates.
Automatically generated - may contain errors
Lab products found in correlation
Micro plus kit, by Qiagen (2 mentions)
Taqman gene expression assay, by Thermo Fisher Scientific (2 mentions)
Rneasy mini, by Qiagen (2 mentions)
Superscript 3 first strand synthesis supermix for qrt pcr kit, by Thermo Fisher Scientific (1 mentions)
Sybr green master mix, by Thermo Fisher Scientific (1 mentions)
Primescript rt master mix kit, by Takara Bio (1 mentions)
Taqman gene expression master mix, by Thermo Fisher Scientific (1 mentions)
Rneasy plus mini kit, by Qiagen (1 mentions)
4 protocols using fast 7900ht
1
Quantitative RNA Expression Analysis
2
Transcriptomic Analysis of Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted with RNEasy Mini or Micro Plus kits (Qiagen) followed by cDNA synthesis using the Superscript III First Strand Synthesis SuperMix for quantitative RT-PCR (qRT-PCR) kit (Life Technologies). Samples were analyzed on an ABI Fast 7900HT under standard conditions using the TaqMan Gene Expression Assays (Life Technologies). Data were normalized to HRPT1 and analyzed using the comparative threshold cycle method.
3
ChIP-qPCR Analysis of Transcription Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The perfused tissues were pooled from 11 pairs of visual cortex (3–4 biological replicates). Each biological replicate constituted to pooled tissue of pups from different litters. They were quenched, lysed and sheared with the Diagenode Bioruptor for 15 cycles of 30-s intervals. Samples were pre-cleared and incubated with MEF2C, HDAC5, and No Antibody, overnight with rotation at 4°C. The immune-complexes were pulled down with magnetic beads, reversed cross-link and purified with phenol–chloroform. Immunoprecipitated chromatin was quantified by real-time quantitative PCR on FAST7900HT (Applied Biosystems) using SYBR-Green master mix (Applied Biosystems), using 10% input. Primers used were: Egr1 forward, 5′-GTGCCCACCACTCTTGGAT-3′, and reverse, 5′-CGAATCGGCCTCTATTTCAA-3′; Arc: forward, 5′-CAGCATAAATAGCCGCTGGT -3′, and reverse, 5′-GAGTGTGGCAGGCTCGTC-3′; Mef2c: forward, 5′-TGCAGAAAAGATTCCCACTTG-3′, and reverse, 5′-AGACACTCACAAGGCAAAGAC -3′. Fold enrichment was calculated by adjusting 10% input to 100% (Ct Input – log210) followed by [Dilution Factor (No Antibody)* (100*2∧ (adjusted input -Ct (IP))] to obtain the fold enrichment.
4
Dacryoadenitis T-cell Phenotyping in Pdcd1−/− Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To characterize helper T-cell phenotype associated with dacryoadenitis in Pdcd1 À/À mice, mRNA levels of IFN-g, Il-4, Il-17A, Il-21, Il-1b, tumor necrosis factor (TNF)-a, Il-6, C-C motif chemokine ligand (Ccl)-3, Ccl-5, Cxcl-9, Cxcl-10, Ccl-20, Ccl-22, and 18S ribosomal RNA as an endogenous control were evaluated. Total RNA was extracted from LGs of female WT and female Pdcd1 À/À mice using an RNA isolation kit (RNeasy Plus Mini kit; Qiagen, Oosterhout, the Netherlands) according to the manufacturer's protocol. RNA was reverse-transcribed to cDNA by using the PrimeScript RT master mix kit (Takara Bio, Shiga, Japan) according to the manufacturer's protocol. Quantitative real-time PCR was performed using specific minor groove binder probes (TaqMan; Applied Biosystems, Foster City, CA) and PCR master mix (TaqMan Gene Expression Master Mix; Applied Biosystems), by a commercial thermocycling real-time PCR system (Fast 7900HT; Applied Biosystems). The primers are described in Table 1. RNA levels were normalized to the level of 18S ribosomal RNA and calculated as DDC t . 38 The mean DC t value of 4month-old WT mice group was used as the calibrator.
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!