Hamster Swab and Tissue cDNA Synthesis

Complementary DNAs (cDNAs) were prepared according to Briese et al. (48 (link)). Briefly, RNA was extracted from hamster swabs and tissues following the QiaAmp Viral RNA extraction protocol (Qiagen), and 11 μl was taken into the SuperScript IV First-Strand cDNA synthesis system (Thermo Fisher Scientific) following the manufacturer’s instructions. After ribonuclease H treatment, second-strand synthesis was performed using Klenow fragment (New England Biolabs) following the manufacturer’s instructions. The resulting double-stranded cDNAs (ds-cDNA) were then purified using Ampure XP bead purification (Beckman Coulter) and eluted into 30 μl of water.

Free full text: Click here

van Doremalen N., Purushotham J.N., Schulz J.E., Holbrook M.G., Bushmaker T., Carmody A., Port J.R., Yinda C.K., Okumura A., Saturday G., Amanat F., Krammer F., Hanley P.W., Smith B.J., Lovaglio J., Anzick S.L., Barbian K., Martens C., Gilbert S.C., Lambe T, & Munster V.J. (2021). Intranasal ChAdOx1 nCoV-19/AZD1222 vaccination reduces viral shedding after SARS-CoV-2 D614G challenge in preclinical models. Science Translational Medicine, 13(607), eabh0755.

Publication 2021

QiaAmp Viral RNA extraction protocol SuperScript IV First-Strand cDNA synthesis system Klenow fragment Ampure XP bead purification

Cdna Hamster Klenow fragment Ribonuclease h Synthesis Tissues Viral rna

Corresponding Organization : National Institutes of Health

Other organizations : Jenner Institute, University of Oxford, Icahn School of Medicine at Mount Sinai

Top 5 similar protocols

Variable analysis

independent variables

RNA extraction method (QiaAmp Viral RNA extraction protocol)
CDNA synthesis method (SuperScript IV First-Strand cDNA synthesis system)
Second-strand synthesis method (Klenow fragment)

dependent variables

Complementary DNAs (cDNAs) prepared from hamster swabs and tissues

control variables

Amount of RNA taken into cDNA synthesis (11 μl)
Ribonuclease H treatment
Double-stranded cDNA (ds-cDNA) purification (Ampure XP bead purification)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!