Difference between revisions of "Part:BBa K1773003:Design"
(→‎Design Notes)
 
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
  Gene mutagenesis was performed using Invitrogen Gene-art multi site mutagenesis PLUS kit.
+
  Gene mutagenesis was performed using Invitrogen GeneArt® Site-Directed Mutagenesis PLUS System kit.
 
These mutagenic primers were used:
 
These mutagenic primers were used:
  

Latest revision as of 15:11, 10 September 2015

I-F type CRISPR-Cas Cas3 gene


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2623
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1344
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2307
    Illegal AgeI site found at 2680
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Gene mutagenesis was performed using Invitrogen GeneArt® Site-Directed Mutagenesis PLUS System kit.

These mutagenic primers were used:

EcoRI mutagenesis:
F: cgttttcaatggcagaatccggcatttgatttggca
R: tgccaaatcaaatgccggattctgccattgaaaacg

XbaI Mutagenesis:
F: cgatcatcattattcttctctggatgctgatgttaatttggg
R: cccaaattaacatcagcatccagagaagaataatgatgatcg

PstI Mutagenesis:
F: gaagactgaactgcctgcggttaaacaacataaac
Re: gtttatgttgtttaaccgcaggcagttcagtcttc


Primers for adding Biobrick prefix and suffix:

Fw: ttaaggaattcgcggccgcttctagatgaatatcttacttgttagccaatgccaaaaaaa

Rev: cggctgcagcggccgctactagtagagatcttcctgccaaaaacccagcgttt