The reliability of transcriptome sequencing was confirmed with qRT–PCR. RNA was extracted from leaves using a TRIzol kit (Accurate Biology, Changsha, China); cDNA was prepared using a reverse transcription kit (Accurate Biology, Changsha, China); and real-time PCR was performed using the SYBR Green method (Accurate Biology, Changsha, China). SiActin (SETIT_026509mg) was used as an internal standard, and relative expression levels were calculated using the 2−ΔΔCt method. At least three replicates were performed in each independent experiment. Primer Premier 5 software (Primer Premier 5.0) was used to design the PCR primers.
Sybr green method
Manufactured by Accurate Biology
Sourced in China
The SYBR Green method is a fluorescent dye-based technique used for quantitative real-time PCR (qPCR) analysis. The SYBR Green dye binds to double-stranded DNA, allowing for the detection and quantification of target DNA sequences during the amplification process. This method provides a simple and cost-effective approach for gene expression analysis, pathogen detection, and other applications requiring real-time DNA quantification.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sybr green method
1
Transcriptome Validation via qRT-PCR
2
Evaluating Transcriptome Sequencing Reliability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To test the reliability of transcriptome sequencing with qRT–PCR, we selected 15 DEGs. RNA was extracted from leaves using a TRIzol kit (Accurate Biology, Changsha, China); cDNA was prepared using a reverse transcription kit (Accurate Biology, Changsha, China); and real-time PCR was performed using the SYBR Green method (Accurate Biology, Changsha, China). SiActin (SETIT_026509mg) was used as an internal standard, and relative expression levels were calculated using the 2−ΔΔCt method [76 (link)]. At least three replicates were performed in each independent experiment. Primer Premier 5 software (Primer Premier 5.0) was used to design the PCR primers (Supplementary Table S1 ).
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!