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Team:University College London/LabBook/Week6
From 2012.igem.org
Contents |
Tuesday 17.7.12
Aim -Testing the competency of the Reinvigorated Cell Line (W3110): After our original attempt to generate competency of the W3100 cell line was inadequate, we undertook a reinvigoration of the W3100 cell line (Expt 5.2). Here we aim to test the competency of the reinvigorated cell line using the same protocol as used in Expt 5.3, by evaluating their growth after transformation with a plasmid of a known high concentration (297ng/ul).
Method
(LOGO) Transformation Protocol 2
Step 1 – Thawing Cells: Use the reinvigorated W3100 cell line created in Week 5 (Expt 5.2)
Step 3 – Addition of BioBrick: To one 2ml eppendorf, add 1ul of pTop plasmid, and to another add nothing – this will be a control.
Step 7 – Adding Broth: SOC media was used, as it is preferred for this protocol.
Step 8 - Incubation: The table below indicates the Ampicillin concentration of the Agar gels.
| Samples | Volume Innoculated | Antibiotic in Gel (conc) | |
|---|---|---|---|
| Plasmid | pTOP | 36ul | Ampicillin (50ug/ml) |
| Control | Positive (No Plasmid) | 36ul | No Antibiotic |
| Negative (No Plasid) | 36ul | Ampicillin (50ug/ml) | |
Wednesday (18.7.12)
Aim - Results from Transformation
Result: Table below indicates there was no growth for our cells, but that the controls worked. Also included is an image of each plate.
| Samples | Growth/No Growth | |
|---|---|---|
| Plasmid | pTOP | No Growth |
| Control | Positive (No Plasmid) | Growth |
| Negative (No Plasid) | No Growth | |
6.2
6.3
