Cdna Circularization
Cdna Circularization - Related Videos
09:44
Rapid Amplification of cDNA Ends
0 Views •
2023
Source: Pablo Sanchez Bosch2, Sean Corcoran2 and Katja Brückner1,2,3 1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research 2Department of Cell and Tissue Biology, 3Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Rapid Amplification of cDNA Ends (RACE) is a technique that allows amplification of full-length cDNA from mRNA by extending to the 3’ or 5’ end, even without prior knowledge of the sequence (Frohman et al.,...
08:39
Quantification of Circular RNAs Using Digital Droplet PCR
0 Views •
Cited by 8 •
2022
This protocol describes the detailed method of digital droplet PCR (dd-PCR) for precise quantification of circular RNA (circRNA) levels in cells using divergent primers.
07:01
Processing the Loblolly Pine PtGen2 cDNA Microarray
0 Views •
Cited by 6 •
2009
The cDNA microarray PtGen2 was developed for gene expression studies in loblolly pine, P. taeda, and other conifer species. Here, we show pre- and post-hybridization handling and washing techniques that can be used with this array to yield better consistency, reduced artifacts, and lower backgrounds.
15:21
RNA Isolation from Embryonic Zebrafish and cDNA Synthesis for Gene Expression Analysis
0 Views •
Cited by 115 •
2009
The isolation of high quality, intact RNA is an essential step in many laboratory protocols. Here, we demonstrate RNA extraction from whole zebrafish embryos and cDNA synthesis for subsequent application in various experimental procedures including gene expression microarray analysis.
02:35
RACE - Rapid Amplification of cDNA Ends
0 Views •
2021
Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer. Since the...
FAQs
Covalent closure changes the molecule’s physical behavior at its ends. Because circular cDNA has no free termini, it can resist degradation by exonucleases more effectively than an open linear form. That increased persistence is important when the molecule must remain intact for downstream copying or sequence-focused analyses of transcript-derived material.
A complementary splint helps align the two cDNA ends in a configuration favorable for ligation. Alternatively, an end-compatible design can provide the matching termini needed for joining. These design choices influence whether the intended junction forms accurately, making end alignment and compatibility central considerations when preparing a circular template for later analysis.
The value of rolling circle amplification is that one circular template can support repeated copying, producing linked copies of the original cDNA sequence. This converts a limited starting molecule into a longer, sequence-linked product, which can increase the transcript-derived material available for sensitive sequencing or related analyses.
A basic workflow begins with linear cDNA, prepares the ends for productive pairing, and brings them together with a complementary splint or an end-compatible design. An enzyme then ligates the aligned ends to create the closed molecule. The resulting product can be carried forward for analyses that benefit from stability or repeated sequence copying.
Researchers may choose this approach when transcript sequences are scarce or when specialized analysis requires multiple linked copies of a cDNA. The method is relevant to transcriptome analysis, isoform characterization, mutation detection, and sensitive sequencing workflows because the circular template combines sequence preservation with an opportunity for repeated copying.
For isoform and mutation studies, the circular template preserves the sequence information present in the original cDNA while enabling linked-copy generation. This can support examination of transcript variants or sequence changes in workflows designed for those purposes. Its usefulness therefore lies not only in molecule stability, but also in preserving information for downstream sequence analysis.