Difference between revisions of "Part:BBa K1789013"
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This is a combination of scaffold and termination.
 
This is a combination of scaffold and termination.
  
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<!-- Add more about the biology of this part here
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===Usage and Biology===
 
===Usage and Biology===
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Usage:
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This part is scaf1 which is associated with a terminator. Scaf1 is used to bind TALE1 and TALE3 protein to make them fixed on the DNA. We create this part to make it more convenient every time we are required to add a scaffold.
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Biology:
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Scaf1 is a DNA scaffold which can bind TALE protein specifically to achieve the fixing of protein or enzyme. This part is scaf1 with a terminator after it.
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==Sequence and Features==
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1789013 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1789013 SequenceAndFeatures</partinfo>
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==Experimental Validation==
  
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This part is validated through four ways: Enzyme cutting, PCR, and Sequence
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===PCR===
  
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<!-- Uncomment this to enable Functional Parameter display
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'''Methods'''
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===Functional Parameters===
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<partinfo>BBa_K1789013 parameters</partinfo>
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<!-- -->
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Usage:
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The PCR is performed with Premix EX Taq by Takara.  
−
This part is scaf1 which is associated with a terminator. Scaf1 is used to bind TALE1 and TALE3 protein to make them fixed on the DNA. We create this part to make it more convenient every time we are required to add a scaffold.
+
  
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The PCR protocol is selected based on the Users Manuel.
 +
The Electrophoresis was performed on a 1％ Agarose glu.
  
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Biology:
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'''Results'''
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Scaf1 is a DNA scaffold which can bind TALE protein specifically to achieve the fixing of protein or enzyme. This part is scaf1 with a terminator after it.
+
 
 +
[[File:1.jpg|150px|]]
 +
 
 +
The result of the agarose electrophoresis was shown on the picture above.
 +
 
 +
===Enzyme cutting===
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 +
'''Methods'''
 +
 
 +
After the assembly ,the plasmid was transferred into the Competent ''E. coli'' DH5α). After culturing overnight in LB,we minipreped the plasmid for cutting.
 +
The preparation of the plasmid was performed with TIANprep Mini Plasmid Kit from ''TIANGEN''. The cutting procedure was performed with EcoRI and XbaI restriction endonuclease bought from ''TAKARA''.
 +
 
 +
The plasmid was cutted in a 20μL system at 37 ℃ for 2 hours.
 +
The Electrophoresis was performed on a 1％ Agarose glu.
 +
 
 +
'''Results'''
 +
 
 +
[[File:SCAF1_Ter酶切.jpg|750px|]]
 +
 
 +
The result of the agarose electrophoresis was shown on the picture above.

Latest revision as of 15:24, 18 September 2015

SCAF1+Ter

This is a combination of scaffold and termination.


Usage and Biology

Usage: This part is scaf1 which is associated with a terminator. Scaf1 is used to bind TALE1 and TALE3 protein to make them fixed on the DNA. We create this part to make it more convenient every time we are required to add a scaffold.


Biology: Scaf1 is a DNA scaffold which can bind TALE protein specifically to achieve the fixing of protein or enzyme. This part is scaf1 with a terminator after it.

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 145
    Illegal AgeI site found at 175
    Illegal AgeI site found at 205
    Illegal AgeI site found at 235
    Illegal AgeI site found at 265
    Illegal AgeI site found at 295
    Illegal AgeI site found at 325
  • 1000
    COMPATIBLE WITH RFC[1000]

Experimental Validation

This part is validated through four ways: Enzyme cutting, PCR, and Sequence

PCR

Methods

The PCR is performed with Premix EX Taq by Takara.

The PCR protocol is selected based on the Users Manuel. The Electrophoresis was performed on a 1％ Agarose glu.

Results

1.jpg

The result of the agarose electrophoresis was shown on the picture above.

Enzyme cutting

Methods

After the assembly ,the plasmid was transferred into the Competent E. coli DH5α). After culturing overnight in LB,we minipreped the plasmid for cutting. The preparation of the plasmid was performed with TIANprep Mini Plasmid Kit from TIANGEN. The cutting procedure was performed with EcoRI and XbaI restriction endonuclease bought from TAKARA.

The plasmid was cutted in a 20μL system at 37 ℃ for 2 hours. The Electrophoresis was performed on a 1％ Agarose glu.

Results

SCAF1 Ter酶切.jpg

The result of the agarose electrophoresis was shown on the picture above.