Difference between revisions of "Part:BBa K4041005"
| Line 3: | Line 3: | ||
<partinfo>BBa_K4041005 short</partinfo> | <partinfo>BBa_K4041005 short</partinfo> | ||
| − | Template for T7 transcription of crRNA for Vibrio pathogenic gene marker. Need to be used with a XbaI overnight clean cut or PCR for the transcription to end at break point, to generate a short crRNA instead of transcription doesn't end. Designed on 100% conserved region. | + | <br>Template for T7 transcription of crRNA for Vibrio pathogenic gene marker. Need to be used with a XbaI overnight clean cut or PCR for the transcription to end at break point, to generate a short crRNA instead of transcription doesn't end. Designed on 100% conserved region. |
https://static.igem.org/mediawiki/parts/0/08/HKIS--crRNA_dem.png | https://static.igem.org/mediawiki/parts/0/08/HKIS--crRNA_dem.png | ||
| − | Need to be used with a T7 promoter based short RNA transcription kit, like MEGAshortscript RNA transcription kit, followed by purification, like spin columns or lithium ion precipitation. | + | <br>Need to be used with a T7 promoter based short RNA transcription kit, like MEGAshortscript RNA transcription kit, followed by purification, like spin columns or lithium ion precipitation. |
| − | + | ||
| + | <br><br> | ||
https://static.igem.org/mediawiki/parts/c/c9/HKIS--ureapage.png | https://static.igem.org/mediawiki/parts/c/c9/HKIS--ureapage.png | ||
| − | 15% UREA Page (Run in room temperature with 80V), indicating that the crRNA is successfully transcribed | + | <br>15% UREA Page (Run in room temperature with 80V), indicating that the crRNA is successfully transcribed |
| − | + | ||
| + | <br><br> | ||
https://static.igem.org/mediawiki/parts/a/a9/HKIS--crRNAbuff.png | https://static.igem.org/mediawiki/parts/a/a9/HKIS--crRNAbuff.png | ||
| − | crRNA used with single stranded template, showcasing that the crRNA best work with 1X NEB2.1 buffer and functions to cleave reporter very rapidly. | + | <br>crRNA used with single stranded template, showcasing that the crRNA best work with 1X NEB2.1 buffer and functions to cleave reporter very rapidly. |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 20:05, 21 October 2021
TDH cas12a crRNA transcription device (crRNA1)
Template for T7 transcription of crRNA for Vibrio pathogenic gene marker. Need to be used with a XbaI overnight clean cut or PCR for the transcription to end at break point, to generate a short crRNA instead of transcription doesn't end. Designed on 100% conserved region.
Need to be used with a T7 promoter based short RNA transcription kit, like MEGAshortscript RNA transcription kit, followed by purification, like spin columns or lithium ion precipitation.
15% UREA Page (Run in room temperature with 80V), indicating that the crRNA is successfully transcribed
crRNA used with single stranded template, showcasing that the crRNA best work with 1X NEB2.1 buffer and functions to cleave reporter very rapidly.
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]