Double-stranded, sequencing

A negative control (a DNA fragment with no sequence change) is used to compare the profiles of the experimental samples. A change in peak appearance or number in the experimental samples indicates a potential variant. This must be further analyzed by double-stranded sequencing to determine the exact nature of the sequence variant. 

The appearance of slow and fast sedimentation peaks at a narrower interval of pH (ApH=0.2) change is the result of PEC particle destruction into the individual macromolecules. The structure of PEC composed of polyampholyte and polyelectrolyte can be represented as double-strand sequences of pairs formed with the help of cooperative systems of ionic and hydrogen bonds. Probably near the lEP, some acidic and basic groups of polyampholytes displaced on loops begin to interact with each other cooperatively with the formation of intramolecular complexes. The mechanism of PEC destruction can be represented as follows ... 

Finally, it is possible to directly sequence double-stranded PCR products. Sequencing DNA from low-melt agarose gels (Kretz, Carson and O Brian 1989) or adding nonionic detergents to the double-stranded DNA prior to sequencing, improves the quality of double-stranded sequencing (R.M. Feldman, this conference). 

MGA has a stem-loop structure containing internal bulged loops [75]. These double-stranded sequences were separated into two single-strand sequences, each of which had no affinity to malachite green, and were tagged to sequences complementary to nucleic acid analytes. This probe is referred to as a binary MGA probe, and provides immediate fluorescent response after hybridization to complementary nucleic acid analytes, thus offering easy and instant detection of specific DNA and RNA [78]. 

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