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Tailed pooled suppression subtractive hybridization (PSSH) adaptors do not alter efficiency.

Robert S Gerrish¹, Steven R Gill

¹Departments of Oral Biology, The State University of New York at Buffalo, NY, USA.

Antonie Van Leeuwenhoek

|June 10, 2010

Summary

This summary is machine-generated.

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Customizing adaptors for Suppression Subtractive Hybridization (SSH) and Pooled Suppression Subtractive Hybridization (PSSH) does not impact subtraction efficiency. This modification allows integration with high-throughput sequencing platforms.

Area of Science:

Genomics
Molecular Biology
Bioinformatics

Background:

Suppression Subtractive Hybridization (SSH) and its derivative, Pooled Suppression Subtractive Hybridization (PSSH), are established methods for analyzing prokaryotic genome structural variations larger than 100 bp.
Traditional SSH analysis relies on Sanger sequencing, limiting compatibility with modern next-generation sequencing (NGS) platforms due to conserved oligonucleotide sequences.

Purpose of the Study:

To investigate whether appending custom sequences to the 5' terminal ends of SSH/PSSH oligonucleotides during nested PCR affects subtraction efficiency.
To determine if this modification enables the use of SSH/PSSH with high-throughput sequencing technologies.

Main Methods:

Comparison of standard PSSH with custom-tailed PSSH using a pool of ten *S. aureus* clinical isolates.

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Assessment of subtraction efficiency between the two PSSH variants.

Main Results:

No statistically significant difference in subtraction efficiency was observed between standard PSSH and custom-tailed PSSH.
The study demonstrates that custom sequences can be appended to adaptors during nested PCR without compromising subtraction efficiency.

Conclusions:

Customizing SSH/PSSH adaptors is feasible and does not reduce subtraction efficiency.
This adaptability is crucial for integrating SSH/PSSH with downstream applications like high-throughput sequencing, broadening its utility in genomic research.