Difference between revisions of "Repeat Midipreps to select new clones for sequencing for failed sequences from Oct. 2"
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*'''Participants:''' Johan, Aaron, Rebecca, Rachel, Allen, Lafia, Menakshi | *'''Participants:''' Johan, Aaron, Rebecca, Rachel, Allen, Lafia, Menakshi (Aaron did the protocol with some help from Johan and Allen) | ||
*'''Date:''' 10/3/2014 | *'''Date:''' 10/3/2014 | ||
*'''Location:''' BioCurious | *'''Location:''' BioCurious | ||
*'''Aims:''' Midiprep colonies | *'''Aims:''' Midiprep colonies from P.FAKS.bovAlphaS1.S (1), P.FAKS.bovAlphaS2(Kex+).S (3), P.FAKS.bov.Beta(A2).S (6) | ||
**(We did not get to midiprep P.FAKS.humkappa(kex-).S (5) tonight because we had to duplicate the glycerol stock) | |||
*'''Materials and Methods'''* | *'''Materials and Methods'''* | ||
**We used the Zyppy Plasmid Midiprep Kit (Protocol = [[Media:D4025i.pdf]]) | **We used the Zyppy Plasmid Midiprep Kit (Protocol = [[Media:D4025i.pdf]]) | ||
***Changes to protocol: Step 9: centrifuge 5 min the 2nd time, not 1 min | |||
**Cultures were grown in 6 ml culture with 6 microliters 50mg/ml carbenicillen from Teknova; they had been innoculated with with single colonies from 9/12/2014 transformation plates; plastic loops (not individually wrapped but from a new bag) were used; cultures were incubated for 24 hours | |||
**Each was done twice to increase the chances of getting the correct sequence because Electra negative control plates had colonies on them, suggesting possible contamination | |||
Revision as of 07:53, 4 October 2014
- Participants: Johan, Aaron, Rebecca, Rachel, Allen, Lafia, Menakshi (Aaron did the protocol with some help from Johan and Allen)
- Date: 10/3/2014
- Location: BioCurious
- Aims: Midiprep colonies from P.FAKS.bovAlphaS1.S (1), P.FAKS.bovAlphaS2(Kex+).S (3), P.FAKS.bov.Beta(A2).S (6)
- (We did not get to midiprep P.FAKS.humkappa(kex-).S (5) tonight because we had to duplicate the glycerol stock)
- Materials and Methods*
- We used the Zyppy Plasmid Midiprep Kit (Protocol = Media:D4025i.pdf)
- Changes to protocol: Step 9: centrifuge 5 min the 2nd time, not 1 min
- Cultures were grown in 6 ml culture with 6 microliters 50mg/ml carbenicillen from Teknova; they had been innoculated with with single colonies from 9/12/2014 transformation plates; plastic loops (not individually wrapped but from a new bag) were used; cultures were incubated for 24 hours
- Each was done twice to increase the chances of getting the correct sequence because Electra negative control plates had colonies on them, suggesting possible contamination
- We used the Zyppy Plasmid Midiprep Kit (Protocol = Media:D4025i.pdf)