Part:BBa_K1698003:Experience

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Applications of BBa_K1698003

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K1698003 was made and tested by Cork iGEM 2015 team:

We had been working with a 62bp Sry detector (K1698002). Some results indicated that this detector showed a degree of non-specificity, giving a positives for female as well as male genomic DNA as target, albeit a lesser number. Given this we thought that a shorter detector might be more specific. We constructed a 32 bp Sry detector. This detector was PCR amplified using primers ordered from IDT and the original template of the 62 bp Sry detector. This detector was first tested with a 32 bp target oligonucleotide and worked extremely well as did a variant with GFP incorporated in the detector plasmid.

Figure Legend: The Sry 32bp detector was tested in either the presence (left) or absence (rgiht) of an oligonucleotide target. The controlled colony count (CCC) is plotted on a the bar chart.

This detector was tested using genomic DNA digested with Pst1 since digest with the six cutter will result in fewer smaller DNA fragments being made that might hybridise on unspecifically when compared to the target being made after the genomic DNA was digested with HaeIII. Further studies were done to investigate digested genomic DNA as a target for both the Sry 62 bp detector and the Sry 32 bp detector. While the results were positive for the male genomic DNA, the presence of some positives with female genomic DNA remained a problem.

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