Difference between revisions of "Part:BBa K2591013:Design"

 
(→‎Source)
Line 13: Line 13:
 
===Source===
 
===Source===
  
−
BBa_K2591000
+
First, we use primers suffix_F(5'-TACTAGTAGCGGCCGCTGCAG-3') and prefix_R(5'-CTCTAGAAGCGGCCGCGAATTC-3') to do a reverse PCR with pSB1C3 backbone, obtaining the linearized backbone. Then we design the primers Prefix_LoxP_bGHpA_F(5'-GAATTCGCGGCCGCTTCTAGAGATAACTTCGTATAGCATACATTATACGAAGTTATTAGAGCTCGCTGATCAGCCTC-3') and Suffix_LoxP_bGHpA_R(5'-CTGCAGCGGCCGCTACTAGTAATAACTTCGTATAATGTATGCTATACGAAGTTATGAGCTCTCCCCAGCATGCCTGCTATTcTC-3') containing LoxP sequence, also, prefix and suffix sequence are added. Using these two primers and BGH polyA containing template to do PCR, we got the LSL part and then used Gibson Assembly to complete the plasmid.
  
 
===References===
 
===References===

Revision as of 07:07, 7 October 2018


LoxP-BGH polyA-LoxP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

BBa_K2591000


Source

First, we use primers suffix_F(5'-TACTAGTAGCGGCCGCTGCAG-3') and prefix_R(5'-CTCTAGAAGCGGCCGCGAATTC-3') to do a reverse PCR with pSB1C3 backbone, obtaining the linearized backbone. Then we design the primers Prefix_LoxP_bGHpA_F(5'-GAATTCGCGGCCGCTTCTAGAGATAACTTCGTATAGCATACATTATACGAAGTTATTAGAGCTCGCTGATCAGCCTC-3') and Suffix_LoxP_bGHpA_R(5'-CTGCAGCGGCCGCTACTAGTAATAACTTCGTATAATGTATGCTATACGAAGTTATGAGCTCTCCCCAGCATGCCTGCTATTcTC-3') containing LoxP sequence, also, prefix and suffix sequence are added. Using these two primers and BGH polyA containing template to do PCR, we got the LSL part and then used Gibson Assembly to complete the plasmid.

References