To investigate the e cient tropism of the recombinant Ad5 vector to CD4-positive cells, shRNA expression was assayed as an indirect index of successful transduction. For this purpose, Total RNA was isolated from cells at 24 hours post-transduction by the mirVana miRNA Isolation Kit (Thermo Fisher, Massachusetts, USA). The rst-strand the complementary DNA (cDNA) was synthesized by stem-loop primers and using reverse transcription enzyme (Thermo Fisher, Massachusetts, USA) according to the kit instructions. The cDNA level was then determined by forward and reverse primers and the SYBR Green Master Mix (YektaTajhiz, Tehran, Iran) as triplicate in the ABI Real-Time PCR system (Applied Biosystems, California, USA). The cDNA expression was normalized with U6-snRNA endogenous control as described previously [38] . The shRNA relative expression was calculated by the 2 -∆Ct [39] (link). HIV-1 proliferation was investigated in culture supernatants harvested 24 hours post-transduction and assayed by p24 antigen ELISA kit.
Reverse transcription enzyme
Manufactured by Thermo Fisher Scientific
Sourced in United States
The reverse transcription enzyme is a critical component used in the process of converting RNA into complementary DNA (cDNA). This enzyme catalyzes the reverse transcription reaction, which is a fundamental step in various molecular biology techniques, including gene expression analysis, viral detection, and RNA sequencing.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using reverse transcription enzyme
1
Evaluating Ad5 Vector Tropism on CD4+ Cells
2
Evaluating Ad5 Vector Tropism on CD4+ Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To investigate the e cient tropism of the recombinant Ad5 vector to CD4-positive cells, shRNA expression was assayed as an indirect index of successful transduction. For this purpose, Total RNA was isolated from cells at 24 hours post-transduction by the mirVana miRNA Isolation Kit (Thermo Fisher, Massachusetts, USA). The rst-strand the complementary DNA (cDNA) was synthesized by stem-loop primers and using reverse transcription enzyme (Thermo Fisher, Massachusetts, USA) according to the kit instructions. The cDNA level was then determined by forward and reverse primers and the SYBR Green Master Mix (YektaTajhiz, Tehran, Iran) as triplicate in the ABI Real-Time PCR system (Applied Biosystems, California, USA). The cDNA expression was normalized with U6-snRNA endogenous control as described previously [38] . The shRNA relative expression was calculated by the 2 -∆Ct [39] (link). HIV-1 proliferation was investigated in culture supernatants harvested 24 hours post-transduction and assayed by p24 antigen ELISA kit.
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!