User:Jose Fabricio Lopez/Notebook/Logbook/2011/07/08

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Objectives

  • Plasmid restriction, with SpeI.


Restriction

  • I need a 2nd restriction for |pUX19-XkR5 plasmid, because the restriction of yesterday shows an unexpected pattern of 3 bands.
  • I cut with SpeI because originally pUX19 has 2 sites, those sites flank insert of RepC-antisenseRNA, with that I expect 2 band:
    • Larger band with 3.9kb
    • Smaller band with 1.4KB(This is because insert for XKR5 plasmid has ~1.4Kb)
Reactives Solutions
SpeI 2μL
Buffer 4 NEB 2μL
BSA10x 2μL
pUX19Extraction 5μL
H2O 9μL
Total 20μL
  • Lane 1= Ruler
  • Lane 2= pUX19XKR5 cut.

pUX19XKR5.SpeI



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