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I am trying to purify a bacterial cytosolic protein from E.coli as heterologous host. It has a C-terminal 6xhis-tag. I have previously purified reasonable yields on NiNTA or Cobalt columns, but...
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Recently, we observed with one particular construct consistently single bp deletions in Gibson assemblies. The 1bp deletions almost exclusively occur in the areas of the primers used to amplify...
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We are frequently using plasmids with a certain backbone. Due to circumstances, some of these were maintained in Ecoli MC1061 and others in DH5a. Recently, we experienced problems with digests...
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I am trying to find out if there are any reasonable guestimates out there on the amount of protein in a total bacterial lysate from a certain amount of cells. All I could find so far is estimates...
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