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Team:Groningen/Notebook/5 September 2009
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===Metal Accumulation=== | ===Metal Accumulation=== | ||
| - | + | '''Colony PCR''' | |
<center>[[Image:Colony_PCR_of_SmtA-1_-_05september2009.jpg|500px]]</center><BR> | <center>[[Image:Colony_PCR_of_SmtA-1_-_05september2009.jpg|500px]]</center><BR> | ||
Colony PCR of [[Team:Groningen/Notebook/24_August_2009#Metal_Accumulation|SmtA colony #1]] using Biobrick VF2/VR primers (lane 1) and RBSforward and SmtAreverse primers (lane 2). Probably a different RBS primer was used, explaining the lack of product. SmtA (including RBS) is 678 bp, PCR product using VF2/VR primers should yield a band of 989 bp. Left and right are both the same 1% agarose (1xTBE) gel on different time points.<BR> | Colony PCR of [[Team:Groningen/Notebook/24_August_2009#Metal_Accumulation|SmtA colony #1]] using Biobrick VF2/VR primers (lane 1) and RBSforward and SmtAreverse primers (lane 2). Probably a different RBS primer was used, explaining the lack of product. SmtA (including RBS) is 678 bp, PCR product using VF2/VR primers should yield a band of 989 bp. Left and right are both the same 1% agarose (1xTBE) gel on different time points.<BR> | ||
| - | + | <BR> | |
| + | '''Restriction analysis''' | ||
| + | <center>[[Image:Restriction_analysis_SmtA_(pSB1AC3)_with_XbaI_%26_PstI-_05september2009.jpg|250px]]</center> | ||
| + | Restriction analysis of of [[Team:Groningen/Notebook/24_August_2009#Metal_Accumulation|SmtA colony #1]] using <i>Pst</i>I and <i>Xba</i>I. Expected fragment was ~700, plasmid was <partinfo>pSB1AC3</partinfo> about 3000 bp. 1% agarose gel (1xTBE) | ||
===Vectors=== | ===Vectors=== | ||
Latest revision as of 15:44, 5 September 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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Wet
GVP Cluster
| E. coli TOP10 Colonies | Name | Plasmid | Date of plating | Antibiotics |
|---|---|---|---|---|
| pCueO-GVP conc. | 4september2009 | Kanamycin/IPTG | ||
| pZntR-GVP conc. | 4september2009 | Kanamycin/IPTG | ||
| pArsR-GVP conc. | 4september2009 | Kanamycin/IPTG | ||
| pLac-GVP conc. | 4september2009 | Kanamycin/IPTG | ||
| (7K) | 4september2009 | Kanamycin/IPTG | ||
| (7K) conc. | 4september2009 | Kanamycin/IPTG | ||
| (7C) | 4september2009 | Kanamycin/IPTG | ||
| (7C) conc. | 4september2009 | Kanamycin/IPTG |
Plates with colonies were stored in fridge, the red indicates a red colony. Plates without colonies were left on the bench.
Transporters
Metal Accumulation
Colony PCR
Colony PCR of SmtA colony #1 using Biobrick VF2/VR primers (lane 1) and RBSforward and SmtAreverse primers (lane 2). Probably a different RBS primer was used, explaining the lack of product. SmtA (including RBS) is 678 bp, PCR product using VF2/VR primers should yield a band of 989 bp. Left and right are both the same 1% agarose (1xTBE) gel on different time points.
Restriction analysis
_with_XbaI_%26_PstI-_05september2009.jpg)
Restriction analysis of of SmtA colony #1 using PstI and XbaI. Expected fragment was ~700, plasmid was about 3000 bp. 1% agarose gel (1xTBE)
Vectors
Dry
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