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Team:Warsaw/Calendar-Stage1/16 July 2010
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(New page: <html> <script type="text/javascript"> ActiveTab='WetLab'; </script> </html> {{TemplateTop}} {{UpperTabs}} {{LowerTabsTeam}} <html> <!-- do not edit above me! --> <br>16.07.2010 Friday ...) |
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<br><br><div class="note">Cloning CommunityRBSes-GFPterminator behind J23100 [promotor]</div> | <br><br><div class="note">Cloning CommunityRBSes-GFPterminator behind J23100 [promotor]</div> | ||
| - | 1. EcoRI/Pst digest of obtained clones to check size of the constructs | + | <br>1. EcoRI/Pst digest of obtained clones to check size of the constructs [Kasia] |
<br>J23100-B0030-GFPterminator sample 1 | <br>J23100-B0030-GFPterminator sample 1 | ||
<br>J23100-B0031-GFPterminator sample 1 | <br>J23100-B0031-GFPterminator sample 1 | ||
| Line 22: | Line 22: | ||
<br>J23100-B0034-GFPterminator sample 1 | <br>J23100-B0034-GFPterminator sample 1 | ||
| - | <br>2. GEL | + | <br>2. GEL [] |
| - | <br>3. Set up liquid culture of ‘cloning intermediates’ communityRBSes-GFPterminator to prepare stocks; samples: | + | <br>3. Set up liquid culture of ‘cloning intermediates’ communityRBSes-GFPterminator to prepare stocks [Cherry]; <br> samples: |
<br>RBS+GFP 10.1 F zag | <br>RBS+GFP 10.1 F zag | ||
<br>RBS+GFP 7.2G zag | <br>RBS+GFP 7.2G zag | ||
| Line 39: | Line 39: | ||
<br>CFP2 | <br>CFP2 | ||
| + | <br><br><div class="note">Checking attendance of internal RBS within mOrange, mCherry, YFP [Kasia, Milena]</div> | ||
| + | <br> Cloning: mOrange after J23100, mCherry after J23100, YFP after J23100 [promotor]: | ||
| + | <br> -> ligation (20ul of fluorescent protein and 2ul of vector with promotor): mOrange + J23100, mCherry + J23100, YFP + J23100; samples were after gel out (from 12.07.2010r), incubation over 1 hour; | ||
| + | <br> -> transformation: 10ul of ligation mix added to competent E. coli DH5alpha cells. | ||
| + | <br> Unfortunately next day there was nothing on the plates after this transformation. | ||
| + | |||
| + | <br><br><div class="note">mCHerry NLS transformation (using E. coli Top10)[Ania P.] -> uzup. </div> | ||
Revision as of 10:48, 31 July 2010
16.07.2010 Friday
Cloning CommunityRBSes-GFPterminator behind J23100 [promotor]
1. EcoRI/Pst digest of obtained clones to check size of the constructs [Kasia]
J23100-B0030-GFPterminator sample 1
J23100-B0031-GFPterminator sample 1
J23100-B0032-GFPterminator sample 1
J23100-B0033-GFPterminator sample 1
J23100-B0034-GFPterminator sample 1
2. GEL []
3. Set up liquid culture of ‘cloning intermediates’ communityRBSes-GFPterminator to prepare stocks [Cherry];
samples:
RBS+GFP 10.1 F zag
RBS+GFP 7.2G zag
RBS+GFP 11.1P zag
RBS+GFP 10.1F
RBS+GFP 11.1P
RBS+GFP 8.1E
RBS+GFP 9.1zag
RBS+GFP 9.1A
RBS+GFP 8.1Ezag
YFP1
YFP2
CFP1
CFP2
Checking attendance of internal RBS within mOrange, mCherry, YFP [Kasia, Milena]
Cloning: mOrange after J23100, mCherry after J23100, YFP after J23100 [promotor]:
-> ligation (20ul of fluorescent protein and 2ul of vector with promotor): mOrange + J23100, mCherry + J23100, YFP + J23100; samples were after gel out (from 12.07.2010r), incubation over 1 hour;
-> transformation: 10ul of ligation mix added to competent E. coli DH5alpha cells.
Unfortunately next day there was nothing on the plates after this transformation.
mCHerry NLS transformation (using E. coli Top10)[Ania P.] -> uzup.
