RT-qPCR was performed with 0.4–1.5 µg total RNA isolated as previously described above. Extracted RNA was annealed with 1875 ng random primers (Invitrogen, Cat#48190-011) for 5 min at 65 °C followed by holding step at 4 °C. Reverse transcription step was performed using 200 U M-MLV reverse transcriptase (Invitrogen, Cat#28025-013) mixed with 30 U RNAse OUT (Invitrogen, Cat#10777-019), M-MLV reverse transcriptase reaction buffer (Invitrogen, Cat#18057-018) and 1 mM final dNTP mix (Invitrogen, Cat#10297-018). Complementary DNA (cDNA) synthesis was carried out at 37 °C for 1 h followed by 95 °C for 5 min cycle and final hold at 4 °C. cDNA was diluted 1:2 or 1:4 in nucleases free water before amplification with LightCycler®480 SYBR Green I Master (Roche, Cat#04-707-516-001) using Light Cycler 480 real-time PCR system (Roche). Absolute abundance of genes relative to B2M housekeeping gene expression was calculated based on a cDNA standard curve using Light Cycler 480 Software 1.7 (Roche). A complete list of primer sequences is provided as supplementary data 4 and 5 .
Final dntp mix
Manufactured by Thermo Fisher Scientific
The Final dNTP mix is a laboratory reagent that contains a balanced mixture of the four deoxyribonucleotides (dATP, dCTP, dGTP, and dTTP) required for various DNA-based applications, such as PCR amplification and DNA sequencing. The product is designed to provide a consistent and reliable source of these essential components for researchers and scientists working in molecular biology and related fields.
Automatically generated - may contain errors
Lab products found in correlation
Rnaseout, by Thermo Fisher Scientific (2 mentions)
Light cycler 480 software 1, by Roche (2 mentions)
M mlv reverse transcriptase, by Thermo Fisher Scientific (2 mentions)
Random primer, by Thermo Fisher Scientific (2 mentions)
Lightcycler 480 sybr green 1 master, by Roche (2 mentions)
Lightcycler 480 real time pcr system, by Roche (2 mentions)
M mlv reverse transcriptase reaction buffer, by Thermo Fisher Scientific (2 mentions)
2 protocols using final dntp mix
1
Quantitative Real-Time PCR Protocol
2
Quantitative RT-PCR for Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RT-qPCR was performed with 0.4 to 1.5 µg total RNA isolated as previously described. Extracted RNA was annealed with 1875 ng random primers (Invitrogen, Cat#48190-011) for 5 min at 65°C followed by holding step at 4°C. Reverse transcription step was performed using 200 U M-MLV reverse transcriptase (Invitrogen, Cat#28025-013) mixed with 30 U RNAse OUT (Invitrogen, Cat#10777-019), M-MLV reverse transcriptase reaction buffer (Invitrogen, Cat#18057-018) and 1 mM final dNTP mix (Invitrogen, Cat#10297-018). Complementary DNA (cDNA) synthesis was carried out at 37°C for 1h followed by 95°C for 5 min cycle and final hold at 4°C. cDNA was diluted 1:2 or 1:4 in nucleases free water before amplification with LightCycler®480 SYBR Green I Master (Roche, Cat#04-707-516-001) using Light Cycler 480 real-time PCR system (Roche). Absolute abundance of genes relative to B2M housekeeping gene expression was calculated based on a cDNA standard curve using Light Cycler 480 Software 1.7 (Roche). A complete list of primer sequences is provided below.
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!