While developing a new hybridization-based molecular test for Cryptosporidium,a protozoan parasite that causes diarrheal illness,a laboratory technician examines the hybridization ability with a closely related,but nontarget,protozoan and also gets a positive test result.In order to improve the stringency of the hybridization reaction for detection of Cryptosporidium only,the technician can do which of the following?
A)Decrease the salt concentration in the hybridization buffer
B)Increase the salt concentration in the hybridization buffer
C)Reduce the reaction temperature during hybridization
D)Remove destabilizing agents,such as formamide,from the hybridization buffer
E)None of the above

Question

Quizplus · Accepted Answer

Stringency in a hybridization assay is affected by the composition of the hybridization buffer,with higher salt concentrations and the presence of destabilizing agents increasing stringency,thereby reducing nontarget hybridizations.In addition,higher temperatures of the hybridization reaction improve stringency by ensuring a higher degree of complementarity between the probe and target nucleic acids,reducing tolerance for nucleotide mismatches that may be present in a closely related,but nontarget sequence.

While Developing a New Hybridization-Based Molecular Test for Cryptosporidium,a Protozoan