Due to water bath failure, the stringent wash step of an SSOP assay occurred at 5°C higher than planned. What is the likely consequence of this deviation?

Performing the stringent wash step of an SSOP (Specific Sequencing Oligonucleotide Probe) assay at a temperature that is 5°C higher than planned is likely to compromise the specificity and efficiency of the assay. The primary purpose of the stringent washing conditions is to reduce non-specific binding and ensure that only tightly bound probes remain associated with the target nucleic acids.

When the temperature is elevated beyond the optimal range, the weaker and potentially non-specific interactions may not be adequately disrupted. As a result, it's possible that some target sequences may not be detected accurately, leading to instances where true positives are missed, resulting in false negative reactions. The elevated temperature could allow the binding of probes that would otherwise be removed under optimal stringent conditions, failing to differentiate between targeted and background signals effectively.

Thus, the likelihood of producing false negatives due to insufficient specificity at an incorrect washing temperature is a significant consequence of this deviation in the protocol.