Difference between revisions of "Part:BBa J176019:Design"

(→‎Design Notes)
(→‎Source)
 
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Reverse: 5'-<u>AAGGCTGCAGCGGCCGCTACTAGT</u>CGGAGGACAGTACTCCGCTC<br>
 
Reverse: 5'-<u>AAGGCTGCAGCGGCCGCTACTAGT</u>CGGAGGACAGTACTCCGCTC<br>
  
−
These primers add a XbaI site upstream of the part and SpeI, NotI, and PstI sites downstream. The PCR amplicon was digested with XbaI/PstI and inserted into an empty V0120 vector.
+
These primers add a XbaI site upstream of the part and SpeI, NotI, and PstI sites downstream. The PCR amplicon was resolved on a gel and the largest band was cut out and purified (the target is repetitive, so special care was taken to ensure the largest product was isolated). The DNA was digested with XbaI/PstI, inserted into an empty V0120 vector, and verified by sequencing.
  
 
===Source===
 
===Source===
  
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TBA
+
Not completely certain, but plasmid pNEBR-X1 (new England Biolabs) is our best guess.
  
 
===References===
 
===References===

Latest revision as of 03:10, 16 October 2011

5xGal4


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

PCR-cloned from the pNEB plasmid, using the following primers:
Forward: 5'-CCTTTCTAGACGGAGTACTGTCCTCCGAGC
Reverse: 5'-AAGGCTGCAGCGGCCGCTACTAGTCGGAGGACAGTACTCCGCTC

These primers add a XbaI site upstream of the part and SpeI, NotI, and PstI sites downstream. The PCR amplicon was resolved on a gel and the largest band was cut out and purified (the target is repetitive, so special care was taken to ensure the largest product was isolated). The DNA was digested with XbaI/PstI, inserted into an empty V0120 vector, and verified by sequencing.

Source

Not completely certain, but plasmid pNEBR-X1 (new England Biolabs) is our best guess.

References