From 2009.igem.org
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| - | Descriptive Title of What You're Doing
| + | Verification Digest of J13002-LuxOD47E-B0015 |
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| | <div class="desc"> | | <div class="desc"> |
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| - | WIKI CODING HERE
| + | *Did a miniprep of overnight cultures and took concentrations. |
| | + | *Set up a Restriction Digest for verification (that the B0015 terminator is present in my contruct) with XbaI and PstI. If successful, we expected to see bands at about 1.7 kb and 3 kb for the contruct (J13002-LuxOD47E-B0015) and the psb1ac3 vector respectively. |
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| | + | Lane 1- J13002-LuxOD47E-B0015 C5, cut with XbaI and PstI |
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| | + | Lane 2- C5 uncut |
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| | + | Lane 3- C6 cut |
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| | + | Lane 4- C6 uncut |
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| | + | Lane 5- C7 cut |
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| | + | Lane 6- C7 uncut |
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| | + | Lane 7- C8 cut |
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| | + | Lane 8- C8 uncut |
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| | + | Lane 9- Size control- J13002-LuxOD47E |
| | + | <Br> |
| | + | Lane 10- Size control- BBK LuxOD47E |
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| | + | *Analysis: From this gel it looks like colonies 5, 6 and 7 may have worked (contain the B0015 terminator) as we see two bands at the expected sizes. Colony 8 has some random bands, so I will ignore this one. Colony 6 looks the most promising as it is the cleanest (no random bands). I will proceed by sending this colony down for sequencing. |
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| | + | *Wrote out a lab blog for this week that I shall post tomorrow morning. |
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Revision as of 20:49, 3 August 2009
University of Calgary
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CAROL
Making Competent Cells
- Prepared sterilized LB broth for growing cells. Followed procedure found in the protocol page for making competent cells.
- After completion of making competent cells, transformed ligated products (pCS26+promoters ligated using NEB Quick Ligase) into XL Gold Ultracompetent cells. These cells were plated on LB+Kan agar plates and was incubated overnight at 37oC.
- As well, as a control, we transformed pBluescript into both XL Gold Ultracompetent cells and Top 10 Cells and plated cells on LB plates to incubate overnight at 37oC.
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CHINMOYEE
Descriptive Title of What You're Doing
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EMILY
Verification Digest of J13002-LuxOD47E-B0015
- Did a miniprep of overnight cultures and took concentrations.
- Set up a Restriction Digest for verification (that the B0015 terminator is present in my contruct) with XbaI and PstI. If successful, we expected to see bands at about 1.7 kb and 3 kb for the contruct (J13002-LuxOD47E-B0015) and the psb1ac3 vector respectively.
Lane 1- J13002-LuxOD47E-B0015 C5, cut with XbaI and PstI
Lane 2- C5 uncut
Lane 3- C6 cut
Lane 4- C6 uncut
Lane 5- C7 cut
Lane 6- C7 uncut
Lane 7- C8 cut
Lane 8- C8 uncut
Lane 9- Size control- J13002-LuxOD47E
Lane 10- Size control- BBK LuxOD47E
- Analysis: From this gel it looks like colonies 5, 6 and 7 may have worked (contain the B0015 terminator) as we see two bands at the expected sizes. Colony 8 has some random bands, so I will ignore this one. Colony 6 looks the most promising as it is the cleanest (no random bands). I will proceed by sending this colony down for sequencing.
- Wrote out a lab blog for this week that I shall post tomorrow morning.
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FAHD
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IMAN
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JAMIE
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JEREMY
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KATIE
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KEVIN
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MANDY
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PATRICK
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PRIMA
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STEFAN
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VICKI
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