Assignments got your hair on fire?

Douse the flames with our full-range writing service!

Experienced academic writing professionals are at your fingertips. Use this handy tool to get a price estimate for your project.

Time course of standard RNA synthesis from three DNA templates "

N2 - All in vitro RNA transcription reactions involve the use of common components: a template, phage RNA polymerase, and ribonucleoside triphosphates (rNTPs). The concentration of rNTPs is a crucial variable that is manipulated to determine the specific activities of labeled RNAs. This protocol describes methods used to synthesize RNAs of low specific activity (trace labeled), medium specific activity, and high specific activity. Also described is how the same principles that determine the level of incorporation of labeled rNTPs are applicable to the synthesis of transcripts containing modified nucleotides, including 5′ cap and internal (body) modifications such as biotinylated uridine, 4-thiouridine, and phosphorothiotated nucleotides. Careful attention to detail in setting up the transcription reaction will permit the synthesis of any number of "designer RNAs.".

AB - All in vitro RNA transcription reactions involve the use of common components: a template, phage RNA polymerase, and ribonucleoside triphosphates (rNTPs). The concentration of rNTPs is a crucial variable that is manipulated to determine the specific activities of labeled RNAs. This protocol describes methods used to synthesize RNAs of low specific activity (trace labeled), medium specific activity, and high specific activity. Also described is how the same principles that determine the level of incorporation of labeled rNTPs are applicable to the synthesis of transcripts containing modified nucleotides, including 5′ cap and internal (body) modifications such as biotinylated uridine, 4-thiouridine, and phosphorothiotated nucleotides. Careful attention to detail in setting up the transcription reaction will permit the synthesis of any number of "designer RNAs.".

Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates.

Synthesis of RNA by In Vitro Transcription | SpringerLink

Using spiked-in synthetic miRNAs, we furtherexamined the two miRNAs with the lowest correlations, and found thedifferences cannot be attributed to differential sensitivity of the twomethods.

Watanabe T, Totoki Y, Toyoda A, Kaneda M,Kuramochi-Miyagawa S, Obata Y, Chiba H, Kohara Y, Kono T, Nakano T,Surani MA, Sakaki Y and Sasaki H: Endogenous siRNAs from naturallyformed dsRNAs regulate transcripts in mouse oocytes. Nature.453:539–543. 2008. : :

Synthesis and Labeling of RNA In Vitro - Current …

Lee RC, Feinbaum RL and Ambros V: The heterochronic gene lin-4 encodes small RNAs withantisense complementarity to lin-14. Cell. 75:843–854. 1993.

Qin XF, An DS, Chen IS and Baltimore D:Inhibiting HIV-1 infection in human T cells by lentiviral-mediateddelivery of small interfering RNA against CCR5. Proc Natl Acad SciUSA. 100:183–188. 2002. : :

Versatile Services that Make Studying Easy
We write effective, thought-provoking essays from scratch
We create erudite academic research papers
We champion seasoned experts for dissertations
We make it our business to construct successful business papers
What if the quality isn’t so great?
Our writers are sourced from experts, and complete an obstacle course of testing to join our brigade. Ours is a top service in the English-speaking world.
How do I know the professor won’t find out?
Everything is confidential. So you know your student paper is wholly yours, we use CopyScape and WriteCheck to guarantee originality (never TurnItIn, which professors patrol).
What if it doesn’t meet my expectations?
Unchanged instructions afford you 10 days to request edits after our agreed due date. With 94% satisfaction, we work until your hair is comfortably cool.
Clients enjoy the breezy experience of working with us
Click to learn our proven method

Synthesis of RNA by in vitro transcription.


In vitro T7 transcription is the synthesis of RNAs using a T7 ..

Novina CD, Murray MF, Dykxhoorn DM,Beresford PJ, Riess J, Lee SK, Collman RG, Lieberman J, Shankar Pand Sharp PA: siRNA-directed inhibition of HIV-1 infection. NatMed. 8:681–686. 2002.

Synthesis and Labeling of RNA In Vitro

Coburn GA and Cullen BR: Potent andspecific inhibition of human immunodeficiency virus type 1replication by RNA interference. J Virol. 76:9225–9231. 2002. : :

Flavivirus RNA synthesis in vitro - ScienceDirect

In contrast to presenting a stepwisedescription of different platforms, we discuss expression profiling ofmature miRNAs by qPCR in four sequential sections: (1) cDNA synthesis;(2) primer design; (3) detection of amplified products; and (4) datanormalization.

Synthesis of RNA by In Vitro Transcription (PDF …

Martínez MA, Gutiérrez A, Armand-Ugón M,Blanco J, Parera M, Gómez J, Clotet B and Esté JA: Suppression ofchemokine receptor expression by RNA interference allows forinhibition of HIV-1 replication. AIDS. 16:2385–2390. 2002.

Basic Protocol 1 describes in vitro synthesis of RNA ..

In conclusion, the short-strand RNA fragments (24 ntlong) of R-Ψ-sgRNA cleaved using the Dicer enzyme exhibitedanti-HIV-1 activity. The sequences of these RNA fragments wereanalyzed to determine the critical sequence for the anti-HIV-1activity. These findings suggest that R-Ψ-sgRNA acts as a miRNA toinhibit HIV-1.

In vitro Assessment of RNA Polymerase I Activity —BIO-PROTOCOL

To test the inhibitory activity of the short-strandRNA fragments on HIV-1 replication in a transient assay, an HIV-1expression plasmid (pNL4-3) and the short-strand RNA fragments weretransfected into HeLa CD4 cells. The virus productionin the culture supernatant was monitored with the HIV-1 p24 antigen(HIV-1 gag gene product) assay. The results of the HIV-1 p24 assayindicated that these RNA fragments had anti-HIV activity andsuppressed HIV-1 replication. Furthermore, our analysis revealedthat the short-strand RNA fragments dose-dependently inhibitedHIV-1 replication ().These results suggest that the short-strand RNA fragments have amiRNA-like effect on the HIV-1 virus.

gRNA synthesis - University of Wisconsin–Madison

We hypothesized that the secondary structure of theR-Ψ-sgRNA had double-stranded regions that may be recognized by theDicer RNase III enzyme, which produces the 24 nt long RNA fragmentsthat are important for RNAi and miRNA. To examine whether the HIV-1suppression mechanism of R-Ψ-sgRNA involved RNAi or miRNA,R-Ψ-sgRNA was synthesized and digested with Dicer(). The size of the digestedRNA fragments was confirmed with 15% polyacrylamide gelelectrophoresis ().

Protocol for synthesis of gRNA for embryonic microinjection

For generating site-specifically and internally radiolabeled RNAs, it usually takes 4 days to finish, provided that the RNA substrate for RNase H cleavage is prepared beforehand and is ready to use. The RNA substrate preparation time is dependent on the type of RNA substrate, from 2 to 3 days for purification of endogenous RNA, to 1 to 2 days for synthesis and purification of in vitro transcribed RNA. The site-specific cleavage of RNA substrate by RNase H (steps 1 to 6 in Basic Protocol 4) can be done on Day 1. The dephosphorylation and re-phosphorylation of the 3′ RNA fragment (steps 7 to 13 in Basic Protocol 4) can be done on Day 2. The ligation of the 5′ RNA fragment to the 3′ RNA fragment (steps 14 to 19 in Basic Protocol 4) can be done on Day 3. The recovery of ligation product (steps 19 to 20 in Basic Protocol 4) can be done on Day 4.

89%
of clients claim significantly improved grades thanks to our work.
98%
of students agree they have more time for other things thanks to us.
Clients Speak
“I didn’t expect I’d be thanking you for actually improving my own writing, but I am. You’re like a second professor!”