In situ probe and inhibitory RNA synthesis using streamlined gene cloning with Gibson assembly

STAR Protocols

Cynthia Wagner¹  Julia Wolf²  Andrew Wolff²  Daniel Lobo² 

[1] Corresponding author;University of Maryland, Baltimore County, Baltimore, MD 21250, USA;

关键词: Developmental biology;    Model Organisms;    Molecular Biology;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: The synthesis of single-stranded riboprobes or double-stranded RNAs for in situ hybridization and gene knockdowns often use vectors that require time-consuming plasmid restriction digests and inefficient gel purifications. Here, we present a faster protocol for the simultaneous plasmid restriction digestion and Gibson assembly of vectors for the synthesis of both riboprobes and double-stranded RNAs for in situ and RNA interference experiments, respectively. We illustrate the protocol with planaria in situ and RNAi assays, but it is applicable to any organism. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

【 授权许可】

Unknown