Difference between revisions of "Part:BBa K1151010"
LeleBiotec (Talk | contribs) (→PCR with VF2 and VR2 primers) |
LeleBiotec (Talk | contribs) |
||
| (3 intermediate revisions by the same user not shown) | |||
| Line 6: | Line 6: | ||
===Boiling prep and digestion with restriction enzymes=== | ===Boiling prep and digestion with restriction enzymes=== | ||
| − | + | Once inserted the construct in a PSB1C3 plasmid (digestion and ligation), we transformed DH5a cells and then we recovered the ligated plasmid by boiling prep. We then proceeded with the digestion with EcoRI and PstI to check. | |
| + | |||
| + | |||
| + | [[File:EPEP1Unisalento.jpg]] | ||
'''Figure 1:''' Digestion result of boiling prep plasmids. | '''Figure 1:''' Digestion result of boiling prep plasmids. | ||
| Line 12: | Line 15: | ||
===PCR with VF2 and VR2 primers=== | ===PCR with VF2 and VR2 primers=== | ||
| − | [[File: | + | [[File:PCR34Unisalento.jpg]] |
'''Figure 2:''' PCR results. | '''Figure 2:''' PCR results. | ||
Latest revision as of 14:54, 2 October 2013
GFP nickel regulated 2
This is a nickel sensitive system driven by the promoter pexbB (BBa_K1151004) with a GFP reporter. Normally the production of GFP is switched on, it is possible to repress the GFP expression using nickel ions in medium and in presence of HpNikR (BBa_K1151000) as trascription factor.
Boiling prep and digestion with restriction enzymes
Once inserted the construct in a PSB1C3 plasmid (digestion and ligation), we transformed DH5a cells and then we recovered the ligated plasmid by boiling prep. We then proceeded with the digestion with EcoRI and PstI to check.
Figure 1: Digestion result of boiling prep plasmids.
PCR with VF2 and VR2 primers
Figure 2: PCR results.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 830