What does SSH mean in UNCLASSIFIED
Suppression Subtractive Hybridisation (SSH) is a molecular biology technique used to identify and isolate specific genes or DNA sequences from a complex mixture. It involves a subtractive hybridisation process that selectively enriches for sequences of interest while suppressing those that are common to both the target and reference samples.
SSH meaning in Unclassified in Miscellaneous
SSH mostly used in an acronym Unclassified in Category Miscellaneous that means Suppression Subtractive Hybridisation
Shorthand: SSH,
Full Form: Suppression Subtractive Hybridisation
For more information of "Suppression Subtractive Hybridisation", see the section below.
Process of SSH
-
RNA Isolation: Total RNA is extracted from two samples, one containing the target genes of interest and the other serving as a reference to subtract common sequences.
-
cDNA Synthesis: Reverse transcription is performed to convert RNA into complementary DNA (cDNA).
-
Adapter Ligation: Adapters (short DNA sequences) are ligated to the ends of both target and reference cDNAs. These adapters allow for specific amplification and hybridisation.
-
Hybridization and Subtraction: The target and reference cDNAs are mixed and hybridised. Perfectly matched duplexes form between common sequences, suppressing them. Non-matching target cDNAs remain single-stranded.
-
PCR Amplification: The single-stranded target cDNAs are selectively amplified using primers specific to the adapters.
-
Cloning and Sequencing: The amplified target fragments are cloned into a plasmid or vector and sequenced to identify the genes or DNA sequences of interest.
Applications of SSH
- Differential gene expression analysis
- Identification of novel genes or transcripts
- Studying gene regulation and cell differentiation
- Diagnostic assays for detecting specific pathogens or genetic mutations
Essential Questions and Answers on Suppression Subtractive Hybridisation in "MISCELLANEOUS»UNFILED"
What is Suppression Subtractive Hybridisation (SSH)?
SSH is a molecular biology technique used to identify and isolate differentially expressed genes between two RNA samples. It involves selectively hybridising and subtracting common sequences, leaving behind unique sequences that represent genes specifically expressed in one sample but not the other.
How does SSH work?
SSH involves three main steps:
- cDNA Synthesis: RNA samples from the two different conditions are converted into cDNA using reverse transcriptase.
- Hybridisation and Subtraction: The cDNA libraries are hybridised together, and common sequences are selectively removed through a series of hybridisation and subtraction steps.
- PCR Amplification: The remaining unique sequences are amplified using PCR, resulting in a subtraction library that contains sequences representing differentially expressed genes.
What is the purpose of SSH?
SSH is primarily used to:
- Identify genes that are specifically expressed or differentially regulated in different cell types, tissues, or developmental stages.
- Study gene expression profiles in response to environmental cues, stimuli, or disease conditions.
- Characterise and compare gene expression patterns between different organisms or experimental conditions.
What are the advantages of SSH?
SSH offers several advantages:
- High Sensitivity: It can detect even low-abundance transcripts by selectively enriching differentially expressed genes.
- Subtracted Hybridization: It eliminates common sequences, allowing for the isolation of unique transcripts.
- Versatility: It can be applied to various RNA sources, including mRNA, miRNA, and lncRNA.
What are the limitations of SSH?
SSH has some limitations:
- Cross-Hybridisation: It may not completely eliminate all common sequences, potentially resulting in false positives.
- Subtraction Bias: The subtraction process can introduce bias, favouring certain transcripts over others.
- Quantity Dependent: It requires sufficient RNA input to obtain reliable results.
Final Words: SSH provides a powerful tool for isolating and characterising specific genes or DNA sequences from complex samples. It enables researchers to study gene expression patterns, identify biomarkers, and investigate biological processes with greater precision.
SSH also stands for: |
|
| All stands for SSH |