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what about other aspects of a biotech/pharma company? Sale, patents, technical support, consultancy, recruitment...?
Companies often search for PhD educated people for these positions.
I met several people who after a PhD or a post-doc left academia and research for a different job in the life science field.
#141176 What else except for research?
Posted
on 11 September 2012 - 08:41 AM
#140906 PCR - consistent false positive results
Posted
on 07 September 2012 - 03:52 AM
1. you can try to digest your plasmid+insert with an enzyme recognizing a site present only in your insert: only one digestion, only one site in the insert (do you have it?). If your plasmid can linearize you can be sure you have your insert in your plasmid.
2. do a PCR with primers that hybridize on your plasmid just after and before your insert. If you have an amplification you have an insert.
3. use a positive positive control (a PCR of your insert before ligation). Just to be sure that the band you see on the gel is your insert. Use 2% gel and a short time migration. Sometimes primers can be visible on the gel and you see them as a kind of band below 200 pb. Do you use a good marker for small fragments?
4. in order to be sure to have 2 inserts in your plasmid, use primers that hybridize on your plasmid (as point 2 so you can have a 240 pb PCR) or digest your plasmid (2 digestions before and after your insert to linearize the plasmid and show a 240 pb fragment (do not cut in inserts)
2. do a PCR with primers that hybridize on your plasmid just after and before your insert. If you have an amplification you have an insert.
3. use a positive positive control (a PCR of your insert before ligation). Just to be sure that the band you see on the gel is your insert. Use 2% gel and a short time migration. Sometimes primers can be visible on the gel and you see them as a kind of band below 200 pb. Do you use a good marker for small fragments?
4. in order to be sure to have 2 inserts in your plasmid, use primers that hybridize on your plasmid (as point 2 so you can have a 240 pb PCR) or digest your plasmid (2 digestions before and after your insert to linearize the plasmid and show a 240 pb fragment (do not cut in inserts)
#140764 protein with good soulbility but doesnt bind to Ni-NTA coloumn
Posted
on 05 September 2012 - 01:57 PM
Yes, probably the tag is hidden by the protein. You can try a C-terminal tag or a longer N-terminal His-tag. How long is your tag now?
Maybe you can try also to lower your imidazole concentration (~20 mM) during the wash step.
Maybe you can try also to lower your imidazole concentration (~20 mM) during the wash step.
