User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/08/17: Difference between revisions
From OpenWetWare
Daniel Catt (talk | contribs) No edit summary |
Daniel Catt (talk | contribs) No edit summary |
||
Line 20: | Line 20: | ||
#* pMAL-pIII in ER2566 + Amp → [[Muratore:Materials/Glycerol stocks | #114]] | #* pMAL-pIII in ER2566 + Amp → [[Muratore:Materials/Glycerol stocks | #114]] | ||
#* pMXB10 in ER2566 + Amp → [[Muratore:Materials/Glycerol stocks | #115]] | #* pMXB10 in ER2566 + Amp → [[Muratore:Materials/Glycerol stocks | #115]] | ||
# Miniprepped | |||
# DNA gel (1% agarose): | # DNA gel (1% agarose): | ||
#* Lane 1: Ladder (10μL) | #* Lane 1: Ladder (10μL) | ||
#* Lane 2: ---- | #* Lane 2: ---- | ||
#* Lane 3: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer A and Pfu Turbo polymerase 1 | #* Lane 3: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer A and Pfu Turbo polymerase 1 [[../16 | 8/16]] | ||
#* Lane 4: ---- | #* Lane 4: ---- | ||
#* Lane 5: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer A and Pfu Turbo polymerase 2 | #* Lane 5: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer A and Pfu Turbo polymerase 2 [[../16 | 8/16]] | ||
#* Lane 6: ---- | #* Lane 6: ---- | ||
#* Lane 7: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer B and Pfu Turbo polymerase 1 | #* Lane 7: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer B and Pfu Turbo polymerase 1 [[../16 | 8/16]] | ||
#* Lane 8: ---- | #* Lane 8: ---- | ||
#* Lane 9: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer B and Pfu Turbo polymerase 2 | #* Lane 9: 5μL test PCR pCMV-SPORT6+BSA with Pfu Turbo buffer B and Pfu Turbo polymerase 2 [[../16 | 8/16]] | ||
#* Lane 10: ---- | #* Lane 10: ---- | ||
#* Lane 11: pMXB10 miniprep from today, 8/17 | #* Lane 11: pMXB10 miniprep from today, 8/17 | ||
#* Lane 12: ---- | #* Lane 12: ---- | ||
#* Lane 13: pMAL-pIII miniprep from today, 8/17 | #* Lane 13: pMAL-pIII miniprep from today, 8/17 | ||
# Innoculated 2 x 500mL of 2xYT media (+ 500μL 100mg/mL ampicillin) with 10mL of the 50mL ER2556 growths from [[../16 | last night]] (pMAL-pIII and pMXB10) | |||
#* After a little under 2 hours, checked the OD @600nm and they were all overgrown by a good bit: | |||
##* pMXB10 #1 = 0.764 | |||
##* pMXB10 #2 = 0.755 | |||
##* pMAL-pIII #1 = 0.663 | |||
##* pMAL-pIII #2 = 0.657 | |||
#* Immediately induced expression with 500μL 1M IPTG in each flask | |||
##* Let grow, shaking overnight @37°C | |||
# Made buffers for protein purification | |||
#* 1M TrisCl pH 8.0, 100mL | |||
##* 12.114g, used HCl to balance to pH 8.0; filtered through 0.45μm membrane | |||
#* 30mM TrisCl pH 8.0 with 20% w/v sucrose, 200mL | |||
##* 6mL of 1M TrisCl stock, 40g of sucrose, balanced pH with HCl; filtered through 0.45μm membrane | |||
==Results== | ==Results== | ||
==Gel== | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | ||
|} | |} |
Revision as of 15:23, 18 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> |
Objective
Bench work
ResultsGel |