By using a Trizol reagent (Life Technologies), total RNA was extracted from cell cultures. Then, reverse transcription was performed with a commercial cDNA kit (Invitrogen, Grand Island, NY, USA). SYBR Green Master Mixes (Invitrogen) was used as a fluorescent dye. Then, qRT-PCR was carried out on One-Step PCR System (Applied Biosystems, Carlsbad, CA, USA). All primer sequences (Sangon Inc., Shanghai, China) are listed in Table S1 .
One step pcr system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The One-Step PCR System is a laboratory instrument designed for performing polymerase chain reaction (PCR) amplification of DNA samples in a single step. It enables the reverse transcription and amplification of RNA samples to be carried out consecutively without the need for additional sample preparation steps.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using one step pcr system
1
RNA Extraction and qRT-PCR Analysis
2
Quantitative Real-Time PCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA extraction and real-time PCR was performed as described in.43 (link) Quantitative PCR of anti-apoptotic markers was carried out using Quantitect primers (Qiagen); all other amplifications were done using TaqMan assay (Applied Biosystems, Rotkreuz, Switzerland). Quantitative PCR was performed using the one-step PCR system (Applied Biosystems). The mean CT was determined from triplicate experiments. miRNA and mRNA levels were normalized to the level obtained for RNU48 and GAPDH, respectively. Changes in expression were calculated using the ΔΔCT method.
3
Quantitative Real-Time PCR of miRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Real-time PCR was performed as described [39 (link)]. Quantitative PCR was carried out using TaqMan assay (Applied Biosystems) and the One-step PCR system (Applied Biosystems). The mean CT was determined from triplicate experiments. miRNA levels were normalized to the level obtained for RNU48. Changes in expression were calculated using the ΔΔCT method.
4
Quantitative real-time PCR protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
By using the Trizol reagent (Life Technologies), total RNA was extracted from cell cultures or fresh tissues. Reverse transcription was performed with a commercial cDNA kit (Takara Bio, Dalian, China). SYBR Green Master Mixes (Takara Bio) was used as a fluorescent dye. qRT-PCR was carried our using One-Step PCR System (Applied Biosystems, Waltham, MA). The primer sequences (AuGCT DNA-SYN Biotech, Beijing, China) are listed in Supplementary Table S2 online.
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!