Team:Exeter/lab book/3gip/wk2

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Revision as of 21:43, 25 September 2012

ExiGEM2012 Lab Book 3GP wk2


Protocols \| Single Gene Plasmids and Enzyme Characterisation \| Showcasing Polysaccharide Production \| The 3-Gene Inducible Plasmid Operon Construction \| Glycobase
9th - 13th July - 16th - 20th July - 23rd - 27th July - 30th July - 3rd August - 6th - 10th August - 13th - 17th August - 20th - 24th August - 27th - 31st August - 3rd - 7th September - 10th - 14th September - 17th - 21st September
	The 3-Gene Inducible Plasmid: 16th - 20th July 2012 Monday 16th July Afternoon • Transformation: RBS Biobrick BBa_B0034, 2012 Distribution Kit Plate 1 2M Tetr_rbs Biobrick BBa_J13002, 2012 Distribution Kit Plate 1 13B Using Invitrogen TOP10 competent cells split into 2 eppendorfs containing 25μl each. Spread on two Ampicillin plates for each transformation using 20μl and 100μls respectively. Tuesday 17th July Afternoon •Adding cultures Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic. Wednesday 18th July Morning The incubator stopped unexpectedly during the night, this meant growth was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were very small and therefore the process was repeated overnight using fresh cultures. Afternoon • Adding cultures Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight. Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic. Thursday 19th July Morning • Mini-Prepping of BBa_B0034 and BBa_J13002 Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with mini-preps. • Gel Electrophoresis on a 2% gel was run to check fragment sizes of BBa_B0034 and BBa_J13002 using the EcoR1 and Pst1 enzymes. BBa_B0034 showed no band but is only 13bp large and BBa_J13002 showed a very faint band.

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 <p><b><u>Wednesday 18th July</u></b></p>
 <p><i><b>Morning</b></i></p>
-<p>The incubator had stopped over night and so growth overnight was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were therefore very small and so the process was repeated overnight using fresh cultures. </p>
+<p>The incubator stopped unexpectedly during the night, this meant growth was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were very small and therefore the process was repeated overnight using fresh cultures. </p>
 <p><i><b>Afternoon</b></i></p>
 <p>•<u> Adding cultures</u></p>
@@ Line 135: / Line 135: @@
 <p>•<u> Mini-Prepping</u> of BBa_B0034 and BBa_J13002</p>
-Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with minipreps. </p>
+Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with mini-preps. </p>
 <p>•<u> Gel Electrophoresis</u> on a 2% gel was run to check fragment sizes of BBa_B0034 and BBa_J13002 using the EcoR1 and Pst1 enzymes. </p>