Difference between revisions of "Part:BBa K1218011:Experience"
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===Characterization of BBa_K1218011=== | ===Characterization of BBa_K1218011=== | ||
| − | SKLBC promotes iGEM in Southern China and found two teams SKLBC-China and SKLBC-GDSYZX in 2015. | + | SKLBC promotes iGEM in Southern China and found two high school software teams SKLBC-China and SKLBC-GDSYZX in 2015. |
As high school team with the BEST SOFTWARE TOOL prize in iGEM 2014 HS, our team continues to focus on developing software to help iGEMers carry on their experiments and focuses more on software. | As high school team with the BEST SOFTWARE TOOL prize in iGEM 2014 HS, our team continues to focus on developing software to help iGEMers carry on their experiments and focuses more on software. | ||
In order to experience synthetic biology, and further promote the impact of iGEM to our schools and Southern China, this summer, we took the experiment to design a new standard BioBrick Part and characterization of an existing part under the instruction of staff from SKLBC. | In order to experience synthetic biology, and further promote the impact of iGEM to our schools and Southern China, this summer, we took the experiment to design a new standard BioBrick Part and characterization of an existing part under the instruction of staff from SKLBC. | ||
During the process, we realized that dry-lab and wet-lab could work close together, to bridge the information gap between science and citizens with educational events. | During the process, we realized that dry-lab and wet-lab could work close together, to bridge the information gap between science and citizens with educational events. | ||
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'''Group''': SKLBC-China 2015 | '''Group''': SKLBC-China 2015 | ||
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'''Group''': SKLBC-GDSYZX 2015 | '''Group''': SKLBC-GDSYZX 2015 | ||
Revision as of 16:36, 18 September 2015
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Characterization of BBa_K1218011
SKLBC promotes iGEM in Southern China and found two high school software teams SKLBC-China and SKLBC-GDSYZX in 2015. As high school team with the BEST SOFTWARE TOOL prize in iGEM 2014 HS, our team continues to focus on developing software to help iGEMers carry on their experiments and focuses more on software. In order to experience synthetic biology, and further promote the impact of iGEM to our schools and Southern China, this summer, we took the experiment to design a new standard BioBrick Part and characterization of an existing part under the instruction of staff from SKLBC. During the process, we realized that dry-lab and wet-lab could work close together, to bridge the information gap between science and citizens with educational events.
Group: SKLBC-China 2015
Author: SKLBC-China 2015
Group: SKLBC-GDSYZX 2015
Author: SKLBC-GDSYZX 2015
Applications of BBa_K1218011
User Reviews
UNIQbe471797bc95ef42-partinfo-00000000-QINU
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CU-Boulder 2014 |
BBa_K1218011 can be targeted to a DNA sequence through the modification of its spacer region.Group: CU-Boulder, 2014 Author: Josephina Hendrix Summary: CU-Boulder demonstrated that BBa_K1218011 can be targeted to a specific DNA sequence through the modification of its spacer region. The original spacer was replaced with one that targeted a neomycin resistance gene. The modified and unmodified plasmids were transformed into cells containing the targeted gene and the decrease in growth with the target sample demonstrates the ability of the spacer to target a specific sequence. Documentation: A 30mer spacer sequence targeting the neomycin phosphotransferase gene was designed and substituted for the original spacer in BBa_K1218011. BW23115 E. coli with the neomycin phosphotransferase inserted into the genome were chemically transformed with the original and modified BBa_K1218011 to compare CRISPR-Cas9 specificity. Transformants were selected for on chloramphenicol. There was a substantial decrease in growth between the non-targeting (1920 colonies) and the targeting sample (8 colonies) that must be accredited to the differences in spacer sequence. As can be seen in Figure 1B), there is growth in the targeting sample. Sequencing showed that all eight colonies had deleted the spacer region and one or both of the adjacent repeats.
Author: Josephina Hendrix Summary: Part BBa_K1218011 was further improved by the addition of the M13 packaging signal (BBa_K1445000) to form the composite part BBa_K1445001. This allows the part to be packaged into M13 phage and delivered to bacteria through phage rather than a transformation method. Documentation:The M13 packaging signal was inserted upstream of the CRISPR-Cas9 part and submitted to the registry as part BBa_K1445001. This new part was packaged into phage and introduced via infection to conjugated neomycin resistant bW23115 E. coli. Sample was plated on chloramphenicol to select for cells infected with the phage delivering the phagemid with the Cas9 part and M13 origin of replication. The growth in figure 2 demonstrates that, with the addition of the M13 origin of replication, BBa_K1218011 can be delivered to cells via recombinant M13 phage. |
UNIQbe471797bc95ef42-partinfo-00000002-QINU