Talk:20.109(S13):Characterize protein expression (Day7): Difference between revisions
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(New page: ==SDS-PAGE analysis tips== You can look up the expected molecular weight of IPC using the sequence file (from Day 1 or Day 5) and [http://www.bioinformatics.org/sms/prot_mw.html this link...) |
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You can look up the expected molecular weight of IPC using the sequence file (from Day 1 or Day 5) and [http://www.bioinformatics.org/sms/prot_mw.html this link] or a similar website. Be sure to add ~ 3 KDa for the size of the N-terminus of pRSET (His tag, etc)! | You can look up the expected molecular weight of IPC using the sequence file (from Day 1 or Day 5) and [http://www.bioinformatics.org/sms/prot_mw.html this link] or a similar website. Be sure to add ~ 3 KDa for the size of the N-terminus of pRSET (His tag, etc)! | ||
If you see two strong bands on your gel, what do you think the second one is? As a starting hint, look back at the Day 6 protocol and carefully consider what is present in the unpurified protein tubes. | If you see two strong bands on your gel, what do you think the second one is? As a starting hint, look back at the Day 6 protocol and carefully consider what is present in the unpurified protein tubes. | ||
What other assays might you consider in relation to SDS-PAGE in order to take a holistic view of your data? | |||
==SDS-PAGE images== | ==SDS-PAGE images== | ||
===T/R=== | |||
[[Image:S13_M2D7_TR_Red.JPG | right |thumb|250px | T/R Gel, Red group -- really W/F Platinum!]] | |||
[[Image:S13_M2D7_TR_Orange.JPG | left |thumb|250px | T/R Gel, Orange group -- really Red]] | |||
[[Image:S13_M2D7_TR_Yellow.JPG | center |thumb|250px | T/R Gel, Yellow group -- really Orange]] | |||
[[Image:S13_M2D7_TR_Green.JPG | right |thumb|250px | T/R Gel, Green group -- really Yellow]] | |||
[[Image:S13_M2D7_TR_Blue.JPG | left |thumb|250px | T/R Gel, Blue group -- really Green]] | |||
[[Image:S13_M2D7_TR_Purple.JPG | center |thumb|250px | T/R Gel, Purple group -- OK]] | |||
[[Image:S13_M2D7_TR_Platinum.JPG | left |thumb|250px | T/R Gel, Platinum group -- OK]] | |||
<br style="clear:both;"/> | |||
===W/F=== | |||
[[Image:S13_M2D7_WF_Red.JPG | right |thumb|250px | W/F Gel, Red group -- really Orange]] | |||
[[Image:S13_M2D7_WF_Orange.JPG | left |thumb|250px | W/F Gel, Orange group -- really Red]] | |||
[[Image:S13_M2D7_WF_Yellow.JPG | center |thumb|250px | W/F Gel, Yellow group -- OK]] | |||
[[Image:S13_M2D7_WF_Green.JPG | right |thumb|250px | W/F Gel, Green group -- OK]] | |||
[[Image:S13_M2D7_WF_Blue.JPG | left |thumb|250px | W/F Gel, Blue group -- OK]] | |||
[[Image:S13_M2D7_WF_Pink.JPG | center |thumb|250px | W/F Gel, Pink group -- really T/R Blue!]] | |||
[[Image:S13_M2D7_WF_Purple.JPG | left |thumb|250px | W/F Gel, Purple group -- really Pink]] | |||
[[Image:S13_M2D7_WF_Platinum.JPG | center |thumb|250px | W/F Gel, Platinum group -- really Purple]] | |||
<br style="clear:both;"/> |
Latest revision as of 14:32, 18 April 2013
SDS-PAGE analysis tips
You can look up the expected molecular weight of IPC using the sequence file (from Day 1 or Day 5) and this link or a similar website. Be sure to add ~ 3 KDa for the size of the N-terminus of pRSET (His tag, etc)!
If you see two strong bands on your gel, what do you think the second one is? As a starting hint, look back at the Day 6 protocol and carefully consider what is present in the unpurified protein tubes.
What other assays might you consider in relation to SDS-PAGE in order to take a holistic view of your data?
SDS-PAGE images
T/R
W/F