User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/24: Difference between revisions
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#* Resuspended pMAL-pIII in 100mL 30mM TrisCl 20% sucrose w/v pH 8.0 | #* Resuspended pMAL-pIII in 100mL 30mM TrisCl 20% sucrose w/v pH 8.0 | ||
#* Resuspended pMXB10 in 25 mL 20mM TrisCl 500mM NaCl pH 8.5 | #* Resuspended pMXB10 in 25 mL 20mM TrisCl 500mM NaCl pH 8.5 | ||
# After gel purification, added 6μL 10x phosphatase buffer, 3μL dH<sub>2</sub>O, and 1μL phosphatase to the 50mL the two plasmids were eluted into | # After gel purification, added 6μL 10x phosphatase buffer, 3μL dH<sub>2</sub>O, and 1μL phosphatase to the 50mL the two plasmids were eluted into. | ||
#* Heat inactivated phosphatase with 5 minutes at 65°C | |||
{| border="1" | |||
|- | |||
!Tube !!sterile H<sub>2</sub>O !!10X ligase buffer !! BSA !!plasmid !!T4 DNA ligase | |||
|- | |||
!pTXB1 + BSA | |||
|13μL || 5μL || 20μL || 10μL || 2μL | |||
|- | |||
!pTXB1 (-) | |||
|33μL || 5μL || 0μL || 10μL || 2μL | |||
|- | |||
!pTXB1.His + BSA | |||
|13μL || 5μL || 20μL || 10μL || 2μL | |||
|- | |||
!pTXB1.His (-) | |||
|33μL || 5μL || 0μL || 10μL || 2μL | |||
|} | |||
==Results== | ==Results== |
Revision as of 14:46, 26 August 2011
AU CHEM-570 Lab Prep | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ObjectiveRepeat BSA cloning into intein vectors with new, correct 3' primer. Bench work
Results
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