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- Member Title Hmmm, I think it's working
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Currently cell biology and developmental biology. Interested in technique development and biochemistry.
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In Topic: media supplements
Yesterday, 08:10 PM
Usually the ATCC sells the supplements required.
In Topic: eGFP vs eGFP-N1
Yesterday, 08:09 PM
Both are plasmids, the N1 construct has the GFP with a MCS at the N-terminus so that the GFP ends up as a C-terminal tag to your protein of interest. The corresponding one for Nterm tag is eGFP-C1.
In Topic: Remove HA Tag in DNA sequence! Help
24 May 2012 - 03:13 PM
Have a close look at the plasmid sequence - the person who cloned the gene may have left restriction site(s) that you can use to either chop out the HA tag or sub clone the gene into a non-HA tagged plasmid.
In Topic: Creating a pGEM-T easy + insert sequence map
23 May 2012 - 01:03 PM
Go here and follow the link on the specifications tab.
In Topic: screening for GPF
23 May 2012 - 01:00 PM
PCR won't tell you if your plasmid has activity, just whether there is plasmid there or not. Restriction digest will/should do the same. Sequencing will also tell you if the gene is there, but activity can only be assessed by looking for the GFP expression either by it fluorescing or on a western blot using anti-GFP antibody.
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