Difference between revisions of "Part:BBa K2371004"
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<partinfo>BBa_K2371004 short</partinfo> | <partinfo>BBa_K2371004 short</partinfo> | ||
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This specific sgRNA generator is for detection of EML4-ALK cancer gene detection in pair with BGIC-Union NT7-dCas9(BBa_K2371000) and CT7-dCas9(BBa_K237101). This part is designed with T7 promotor and T7 terminator and can effectively generate specific RNA in vitro. The product can have RNA concentration of 1500 ng/ul or higher with MEGAscript™ T7 Transcription Kit and after the purification and collection using Ambion® mirVana™ miRNA Isolation Kit. | This specific sgRNA generator is for detection of EML4-ALK cancer gene detection in pair with BGIC-Union NT7-dCas9(BBa_K2371000) and CT7-dCas9(BBa_K237101). This part is designed with T7 promotor and T7 terminator and can effectively generate specific RNA in vitro. The product can have RNA concentration of 1500 ng/ul or higher with MEGAscript™ T7 Transcription Kit and after the purification and collection using Ambion® mirVana™ miRNA Isolation Kit. | ||
[[File:BGIC-Union Part sgRNA Figure1 plasmid.png|700px]] | [[File:BGIC-Union Part sgRNA Figure1 plasmid.png|700px]] | ||
| − | < | + | <h4>Figure 1. The plasmid of sgRNA generator. The sgRNA generator Biobrick is composed of a T7 promotor, coding sequence for sgRNA and a T7 terminator.</h4> |
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| + | The sequence of enzyme sites, T7 promotor, sgRNA sequence and T7 terminator were synthesized by SOEPCR designed by DNAworks. The gel electrophoresis of the second amplification in SOEPCR using the oligo DNA in the start and the end of the sequence as primers validate the success of the synthesis. | ||
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| + | [[File:BGIC-Union Part sgRNA Figure2.png|700px]] | ||
| + | <h4>Figure 2. The electrophoresis of second amplification using the using the oligo DNA in the start and the end of the sequence as primers.The desired length should be around 200bp.</h4> | ||
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Revision as of 07:11, 31 October 2017
sgRNA generator for EML4-ALK variant A 23
This specific sgRNA generator is for detection of EML4-ALK cancer gene detection in pair with BGIC-Union NT7-dCas9(BBa_K2371000) and CT7-dCas9(BBa_K237101). This part is designed with T7 promotor and T7 terminator and can effectively generate specific RNA in vitro. The product can have RNA concentration of 1500 ng/ul or higher with MEGAscript™ T7 Transcription Kit and after the purification and collection using Ambion® mirVana™ miRNA Isolation Kit.
Figure 1. The plasmid of sgRNA generator. The sgRNA generator Biobrick is composed of a T7 promotor, coding sequence for sgRNA and a T7 terminator.
The sequence of enzyme sites, T7 promotor, sgRNA sequence and T7 terminator were synthesized by SOEPCR designed by DNAworks. The gel electrophoresis of the second amplification in SOEPCR using the oligo DNA in the start and the end of the sequence as primers validate the success of the synthesis.
Figure 2. The electrophoresis of second amplification using the using the oligo DNA in the start and the end of the sequence as primers.The desired length should be around 200bp.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]