One such technique described as genetic bit analysis (GBA) is based on hybridization capture of a single stranded PCR product to a sequence-specific microplate-bound primer,
followed by enzyme-mediated single base extension of the primer across the polymorphic site, enabling direct determination of SNP through colorimetry.
The technique has been used as a diagnostic tool in human paternity tests as well as in pedigree analysis.
Among plant systems, the successful use of the technique has been demonstrated in onions for distinguishing between plastomes of cytoplasmic male sterile (CMS) and fertile lines, which differ by a single SNP.
It has been shown that semi-automated GBA can be conducted using 96 well micro titer plates.
Normal Cytoplasm
| 1. single-strand obtained from PCR Product |
2. Single-strand hybridized to GBA primer |
| 3. Incorporation of Fluorescein-labelled ddG |
4. Colorimetric detection (yellow colour) |
Sterile Cytoplasm
| 1. Single-strand obtained from PCR product |
2. Single strand Hybridized to GBA primer |
| 3. incorporation of biotin labelled ddT |
4. colorimetric detection (blue colour) |
|
