Rrna Removal
Rrna Removal - Related Videos
10:57
16S rRNA Sequencing: A PCR-based Technique to Identify Bacterial Species
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2023
Source: Ewa Bukowska-Faniband1, Tilde Andersson1, Rolf Lood1 1 Department of Clinical Sciences Lund, Division of Infection Medicine, Biomedical Center, Lund University, 221 00 Lund, Sweden Planet Earth is a habitat for millions of bacterial species, each of which has specific characteristics. Identification of bacterial species is widely used in microbial ecology to determine biodiversity of environmental samples and medical microbiology to diagnose infected patients. Bacteria can be classified...
12:37
Efficient Nucleic Acid Extraction and 16S rRNA Gene Sequencing for Bacterial Community Characterization
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Cited by 42 •
2016
We describe an efficient, robust, and cost effective method for extracting nucleic acid from swabs for characterization of bacterial communities using 16S rRNA gene amplicon sequencing. The method allows for a common processing approach for multiple sample types and accommodates a number of downstream analytic processes.
09:12
DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
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2019
Here we present a protocol for DNAzyme-dependent cleavage of RNA. This enables fast and site-dependent analysis of RNA 2’-O-methylation. This approach can be used for the preliminary or major assessment of snoRNA activity.
06:43
Rotary Evaporation to Remove Solvent
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2023
Source: Dr. Melanie Pribisko Yen and Grace Tang — California Institute of Technology Rotary evaporation is a technique most commonly used in organic chemistry to remove a solvent from a higher-boiling point compound of interest. The rotary evaporator, or "rotovap", was invented in 1950 by the chemist Lyman C. Craig. The primary use of a rotovap is to dry and purify samples for downstream applications. Its speed and ability to handle large volumes of solvent make rotary evaporation a preferred...
03:02
Nictitating Membrane Removal: A Surgical Procedure to Remove Nictitating Membrane from Rabbit Eye
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2025
In this video, we demonstrate a surgical procedure to remove nictitating membrane from rabbit’s eye.
FAQs
The key selectivity comes from sequence complementarity: oligonucleotide probes are designed to hybridize with abundant rRNA molecules in the total RNA mixture. Once those hybrids form, RNase H digestion can target the RNA paired with the probe, or probe-based capture can remove the hybridized material. The remaining sample therefore contains a higher relative proportion of non-ribosomal transcripts.
Probe-based capture and RNase H digestion remove rRNA through different routes. Digestion uses the probe-rRNA hybrid as a substrate for enzymatic cleavage, whereas capture retains the hybridized rRNA so it can be separated from the rest of the sample. Both strategies pursue the same enrichment goal, but they represent distinct ways to deplete the dominant RNA class.
Compared with poly(A) selection, rRNA removal does not rely solely on that selection strategy. This distinction matters when the experiment includes messenger RNA together with other coding or noncoding transcripts. By reducing the space occupied by abundant rRNA, the method can allocate more sequencing reads to low-abundance targets and support analyses of transcript structure and gene expression.
The immediate outcome is relative enrichment rather than creation of new transcripts. Messenger RNA and other transcripts become more represented after rRNA reduction, which can improve the usefulness of sequencing data for examining low-abundance coding and noncoding RNAs. Researchers can also investigate transcript structure in a sample that is not restricted to poly(A)-selected molecules.
Starting with total RNA, the workflow introduces complementary oligonucleotide probes that hybridize to the targeted abundant rRNA molecules. The sample then follows one of two routes: RNase H digestion of probe-associated RNA or probe-based capture and separation. The resulting RNA fraction is used for sequencing, with more reads available for non-ribosomal transcripts.
It is particularly useful when researchers want sequencing access to low-abundance coding and noncoding RNAs, transcript structure, or gene expression without relying exclusively on poly(A) selection. The approach is therefore suited to transcriptomics questions requiring a broader representation of RNA species than a strategy centered only on poly(A)-selected material.
Successful depletion reduces the proportion of sequencing data dominated by abundant ribosomal molecules and increases the relative representation of messenger RNA and other transcripts. This can improve sequencing efficiency and read allocation, allowing the dataset to devote more attention to targets that may otherwise be underrepresented, including low-abundance coding and noncoding RNAs.
In biology, the method connects RNA sample preparation with questions about how genes are expressed and how transcripts are structured. Removing a highly abundant RNA class before sequencing helps researchers examine messenger RNA alongside other coding and noncoding transcripts. That broader transcript view is valuable when the study’s biological signal is not confined to poly(A)-selected RNA.