Team:BCCS-Bristol/Calendar-Notebook/15 July 2008

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Revision as of 09:56, 6 August 2008

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BioBrick Transformation

SOC medium was made in preparation for Biobrick transformation. This consisted of

Component	Amount
Trypton	2g
Yeast Extract	0.5g
NaCl	0.05g

in 95ml of water, then autoclaved. After this the following was added:

Component	Amount
MgCl₂ (2 M)	0.5ml
Glucose (1 M)	2ml

The medium is stored at -20^oC. This solution is from Molecular Cloning 3 A.2 SOB/SOC Medium.

Chambers for bacteria-moving-bead assay

The first trial in making wells as in the diagram (See below) was made, but it was found that a different glue is needed. Nevertheless, in the first chambers a blue dye in 0.3% agar (to represent aspartate) and an orange dye in 0.1% agar to represent the bacteria was put (beads were added in later experiments). Pictures were taken at 30 minutes intervals. This was to see how long a chemotatic gradient would take to set up. However, after three hours no gradient was obvious, so the chambers were left overnight.

Comparision of motility between E. coli MG1655 and MC1000

The motility of both strains was tested under different conditions:

- agar concentration: 0.3 % and 0.5 %

- incubation temperatures: 25^oC and 30^oC

The OD₆₀₀ was adjusted to 1.25 and the bacteria were inoculated in the middle of an LB-plate without yeast extract, but with streptomycin. After an incubation overnight there was no growth of MG1655 visible, because only MC1000 is resistant. Therefore, the experiment should be repeated.

@@ Line 15: / Line 15: @@
 </div>
-Made SOC medium in preparation for Biobrick transformation this consisted of
+==BioBrick Transformation==
+SOC medium was made in preparation for Biobrick transformation. This consisted of
 {| {{table}}
-| align="center" style="background:#f0f0f0;"|'''Trypton'''
+| align="center" style="background:#f0f0f0;"|'''Component'''
-| align="center" style="background:#f0f0f0;"|'''2g'''
+| align="center" style="background:#f0f0f0;"|'''Amount'''
+|-
+| Trypton||2g
 |-
 | Yeast Extract||0.5g
@@ Line 28: / Line 32: @@
 |}
-in 95ml of water, then autoclaved. after this add this:
+in 95ml of water, then autoclaved. After this the following was added:
 {| {{table}}
-| align="center" style="background:#f0f0f0;"|'''MgCl2'''
+| align="center" style="background:#f0f0f0;"|'''Component'''
-| align="center" style="background:#f0f0f0;"|'''0.5ml 2M solution'''
+| align="center" style="background:#f0f0f0;"|'''Amount'''
 |-
-| Glucose||2ml 1M
+| MgCl<sub>2</sub> (2 M)||0.5ml
+|-
+| Glucose (1 M)||2ml
 |-
 |
 |}
-then freeze at -20<sup>o</sup>C. this solution is from Molecular Cloning 3 A.2 SOB/SOC Medium.
+The medium is stored at -20<sup>o</sup>C. This solution is from Molecular Cloning 3 A.2 SOB/SOC Medium.
+==Chambers for bacteria-moving-bead assay==
-had a go at making wells as in the diagram  (See below), needs diff glue. then put a blue dye in 0.3% agar (to represent aspartate) and an orange dye in 0.1% agar to represent the bacteria. took pictures at 30minute intervals. this was to see how long a chemotatic gradient would take to set up, however after 3 hours no gradient was obious so left iovernight.
+The first trial in making wells as in the diagram (See below) was made, but it was found that a different glue is needed. Nevertheless, in the first chambers a blue dye in 0.3% agar (to represent aspartate) and an orange dye in 0.1% agar to represent the bacteria was put (beads were added in later experiments). Pictures were taken at 30 minutes intervals. This was to see how long a chemotatic gradient would take to set up. However, after three hours no gradient was obvious, so the chambers were left overnight.
-Recorded motility comparison, turns out we messed up and tried to grow MG1655 on streptomycin, therefore we made a repeat of this experimen
 [[Image:BCCS_Bristol-Wetlab-Well_diagram.JPG | 800px]]
+==Comparision of motility between ''E. coli'' MG1655 and MC1000==
+The motility of both strains was tested under different conditions:
+- agar concentration: 0.3 % and 0.5 %
+- incubation temperatures: 25<sup>o</sup>C and 30<sup>o</sup>C
+The OD<sub>600</sub> was adjusted to 1.25 and the bacteria were inoculated in the middle of an LB-plate without yeast extract, but with streptomycin. After an incubation overnight there was no growth of MG1655 visible, because only MC1000 is resistant. Therefore, the experiment should be repeated.