Difference between revisions of "Part:BBa K2440000"
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Locker A is a miRNA locker with two identical functional modules to target miR-214. | Locker A is a miRNA locker with two identical functional modules to target miR-214. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | It is one of the experimental cases which were constructed to examine the scalability, reliability and efficiency of the Overlapped Oligo Assembly (OOA) method. | |
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| + | ===Overlapped Oligo Assembly method=== | ||
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| + | In OOA method, modularized DNA parts is designed into the chemically synthesized DNA oligo, and overlapped 5’-overhangs are added for assembly. After that, dsDNAs can be obtained by annealing of two complementary single-strand oligonucleotides, then phosphorylated by T4 Polynucleotide Kinase to facilitate the following ligation reaction. T4 DNA ligase is use for their ligation onto pSB1C3 plasmid backbone. High-throughput one-step assemble of multiple short dsDNA nucleotides can therefore be achieved, and the correct assembling order is guaranteed by using identical overlap sequences among oligos, serving as zipcodes. The zipcodes are introduced as the intervals in a miRNA Locker. | ||
| + | The dsDNA assembles are capable of being sub-cloned into plasmid backbones for amplification and long-term storage. In view of that, a prefix and a suffix is added to the dsDNA oligos forming the 5’ and 3’ terminus of the assembly, ensuring that the plasmids we constructed are compatible with BioBrick RFC[10]. | ||
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| + | ===Sequence and Features=== | ||
| + | |||
<partinfo>BBa_K2440000 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2440000 SequenceAndFeatures</partinfo> | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2440000 parameters</partinfo> | <partinfo>BBa_K2440000 parameters</partinfo> | ||
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Revision as of 18:42, 1 November 2017
Locker A
Locker A is a miRNA locker with two identical functional modules to target miR-214.
Usage and Biology
It is one of the experimental cases which were constructed to examine the scalability, reliability and efficiency of the Overlapped Oligo Assembly (OOA) method.
Overlapped Oligo Assembly method
In OOA method, modularized DNA parts is designed into the chemically synthesized DNA oligo, and overlapped 5’-overhangs are added for assembly. After that, dsDNAs can be obtained by annealing of two complementary single-strand oligonucleotides, then phosphorylated by T4 Polynucleotide Kinase to facilitate the following ligation reaction. T4 DNA ligase is use for their ligation onto pSB1C3 plasmid backbone. High-throughput one-step assemble of multiple short dsDNA nucleotides can therefore be achieved, and the correct assembling order is guaranteed by using identical overlap sequences among oligos, serving as zipcodes. The zipcodes are introduced as the intervals in a miRNA Locker. The dsDNA assembles are capable of being sub-cloned into plasmid backbones for amplification and long-term storage. In view of that, a prefix and a suffix is added to the dsDNA oligos forming the 5’ and 3’ terminus of the assembly, ensuring that the plasmids we constructed are compatible with BioBrick RFC[10].
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]