Total RNAs were isolated using the miRNeasy Mini Kit (Qiagen, Germany). miRNAs were transcribed into cDNAs using the Mir-X miRNA First Strand Synthesis kit (#RR638315, TaKaRa), while mRNAs and lncRNAs were transcribed into cDNAs using the HiScript II RT kit (Vazyme, Nanjing, China). Real-time PCR was detected under the StepOnePlus platform (Applied Biosystems) using the HiScript II One Step RT-PCR SYBR Green Kit (Vazyme). The relative expression levels of the targeted genes were normalized to β-actin, GAPDH, and U6, and then calculated using the 2−ΔΔCt method. The cytoplasmic and nuclear RNAs were isolated using the Cytoplasmic & Nuclear RNA Purification Kit (Norgen Biotek Corp). The RNAs were then transcribed into cDNAs and detected using the above methods. GADPH and U3 were used as cytoplasm-located and nucleus-located markers separately. The primers used in the PCR were described in Supplementary Tables S3 and S4 .
Hiscript 2 one step rt pcr sybr green kit
Manufactured by Vazyme
Sourced in China
The HiScript II One Step RT-PCR SYBR Green Kit is a reagent kit designed for reverse transcription and real-time PCR amplification using SYBR Green detection. It enables the conversion of RNA to cDNA and subsequent real-time PCR in a single reaction.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using hiscript 2 one step rt pcr sybr green kit
1
Isolation and Detection of RNAs
2
Quantifying Plant Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from leaves using the RNAprep Pure Plant Plus Kit (spin column) (Takara, Dalian, China) based on specifications. Approximately 1 μg total RNA was reverse-transcribed into cDNA using the “StarScript ǁ First-stand cDNA Synthesis Mix with gDNA Remover”. The qRT-PCR was performed on using a Roche LightCycler 480II detection system, using Hiscript II one-step RT-PCR SYBR Green kit (Vazyme, Nanjing, China). The primers used in this study are listed in Supplementary Table 1 . Each sample was conducted in three biological replications. CnActin (KF305683) was used as the reference gene. The relative expression level of each gene was calculated as 2-ΔΔCT equation [85 (link)].
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!