IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/09/06

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(Floor Two)
Current revision (13:26, 30 September 2013) (view source)
(Floor One)
 
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==Floor One==
==Floor One==
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[[Image:GEL8.png|thumb| Fig 1. Gel image showing the PCR from the 16S sequences. Lane 1 is a 1Kb+ ladder, and lanes 2-20 alternatively contain the 10 samples. Only lanes 6 and 18 didn't work.]]
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[[Image:GEL5.png|thumb| Fig 2. Gel image showing the GUS product in lanes 2-5, and a 1Kb+ ladder in lane 1.]]
2x 1% Agarose gels made, one 100ml and one 60ml.
2x 1% Agarose gels made, one 100ml and one 60ml.
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Loaded the 16S sequenced PCRs in the 100ml gel with a 1Kb+ ladder, and run at 110mV for 50 mintues.  
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Loaded the 16S sequenced PCRs in the 100ml gel with a 1Kb+ ladder, and run at 110mV for 50 mintues ''fig 1''.  
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Loaded the GUS PCR in the 60ml gel, and run for 50 minutes at 110mV.  
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Loaded the GUS PCR in the 60ml gel, and run for 50 minutes at 110mV ''fig 2''.  
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4x 500ml SFM made.
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4x 500ml SFM made. More streak purifications and lawns carried out.
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Both gels ran and showed enough product to gel extract.  
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Of the 10 16S lanes, 8 ran, and GUS showed enough product to gel extract, using the QIAqick kit. Excised fragments from the gels were dissolved in Buffer QG, incubated and centrifuged through a spin colum with isopropanol at 13,000rpm for one minute. This was washed with Buffer PE and eluted in sterile water, and stored in the freezer.
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The 16S PCR products will be prepped and sent off for sequencing.
==Floor Two==
==Floor Two==

Current revision

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Floor One

Fig 1. Gel image showing the PCR from the 16S sequences. Lane 1 is a 1Kb+ ladder, and lanes 2-20 alternatively contain the 10 samples. Only lanes 6 and 18 didn't work.
Fig 1. Gel image showing the PCR from the 16S sequences. Lane 1 is a 1Kb+ ladder, and lanes 2-20 alternatively contain the 10 samples. Only lanes 6 and 18 didn't work.
Fig 2. Gel image showing the GUS product in lanes 2-5, and a 1Kb+ ladder in lane 1.
Fig 2. Gel image showing the GUS product in lanes 2-5, and a 1Kb+ ladder in lane 1.

2x 1% Agarose gels made, one 100ml and one 60ml. Loaded the 16S sequenced PCRs in the 100ml gel with a 1Kb+ ladder, and run at 110mV for 50 mintues fig 1.

Loaded the GUS PCR in the 60ml gel, and run for 50 minutes at 110mV fig 2.

4x 500ml SFM made. More streak purifications and lawns carried out.

Of the 10 16S lanes, 8 ran, and GUS showed enough product to gel extract, using the QIAqick kit. Excised fragments from the gels were dissolved in Buffer QG, incubated and centrifuged through a spin colum with isopropanol at 13,000rpm for one minute. This was washed with Buffer PE and eluted in sterile water, and stored in the freezer.

The 16S PCR products will be prepped and sent off for sequencing.

Floor Two

The biobrick K1041000 was compared to Bba_J04450 by performing a restriction digest of both with NdeI and EcoRI+PstI and running the samples on an Agarose gel.

Analysis of Restriction enzyme digest. Lane 1-3 contains uncut K1041000, K1041000 digested with NdeI then EcoRI and PstI Respectively. Lanes 4-6 contains uncut J04450. J04450 digested with NdeI then EcoRI and PstI respectively.
Analysis of Restriction enzyme digest. Lane 1-3 contains uncut K1041000, K1041000 digested with NdeI then EcoRI and PstI Respectively. Lanes 4-6 contains uncut J04450. J04450 digested with NdeI then EcoRI and PstI respectively.



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