In most samples, total RNA was prepared using TRIzol Reagent (Ambion, 15596018, TX, USA). Reverse transcription-PCR was carried out using a High Capacity cDNA Reverse Transcription kit (Applied Biosystems, 4368814, MA, USA). For total RNA purification from OVA-tetramer+ CD8+ T cells, CellAmp® Whole Transcriptome Amplification Kit (Real Time) Ver.2 (Takara, 3734, Shiga, Japan) was used according to the manufacturer’s instructions. Real-time PCR was performed using a Step One real-time PCR system (Applied Biosystems, 4368813). Sequences of primers in this study are shown in Additional file 1 : Table S1. Levels of target mRNAs were normalized to Gapdh and fold-induction of transcripts was calculated using the ddCT method.
Cellamp whole transcriptome amplification kit real time ver 2
Manufactured by Takara Bio
Sourced in Japan
The CellAmp® Whole Transcriptome Amplification Kit (Real Time) Ver.2 is a product designed for whole transcriptome amplification from a small number of cells or single cells. The kit enables amplification of the entire transcriptome in a quick and efficient manner, providing a comprehensive representation of gene expression profiles.
Automatically generated - may contain errors
2 protocols using cellamp whole transcriptome amplification kit real time ver 2
1
RNA Isolation and qRT-PCR Analysis
2
Quantification of Immune Transcription Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Purified genomic DNA was isolated from DCs using the CellAmp™ Whole Transcriptome Amplification kit (Real Time) Ver2 (Code No. 3734, TaKara). Relative quantitative reverse transcriptase polymerase chain reaction (RT‐PCR) was performed to detect the expression of NF‐κb, JNK, IFN‐γ, and GAPDH with SYBR Green PCR kit (Takara, Dalian, China) using the StepOnePlus™ Real‐Time PCR System (Life Technoligies, Carlsbad, NM, USA). The specific RT‐PCR primers are shown in Table 1 . The following PCR amplification protocol was used: (1) an initial denaturation at 95°C for 30 s, (2) 40 cycles of 95°C for 5 s and 60°C for 34 s, and (3) a final extension at 95°C for 15 s, 60°C for 1 min, and 95°C for 15 min. The expression changes of genes were calculated using the △Ct method with GAPDH as internal control.
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!