Team:BYU Provo/Notebook/SmallPhage/Summerexp/Period2/Exp/7.27 Bacteria Growth Curve After Dilution
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<font color="#333399" size="3" font face="Calibri"> | <font color="#333399" size="3" font face="Calibri"> | ||
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+ | : <u> '''Small Phage''' </u> </font> | ||
: [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]] | : [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]] | ||
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'''I) Purpose''' | '''I) Purpose''' | ||
* Determine the growth curve of bacteria after dilution. This information will be used to estimate the amount of bacterial growth during incubation in mutagenesis experiments. | * Determine the growth curve of bacteria after dilution. This information will be used to estimate the amount of bacterial growth during incubation in mutagenesis experiments. | ||
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'''II) Expected Outcome''' | '''II) Expected Outcome''' | ||
* A sigmoidal growth curve for each dilution. | * A sigmoidal growth curve for each dilution. | ||
* The bigger the initial dilution factor, the longer it takes for bacteria to reach its growth plateau. | * The bigger the initial dilution factor, the longer it takes for bacteria to reach its growth plateau. | ||
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'''III) Reagants Used''' | '''III) Reagants Used''' | ||
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* Pure LB was used to as blank | * Pure LB was used to as blank | ||
* 1mL of E coli solution at various concentration and time points was pipetted into each cuvette for measurement. | * 1mL of E coli solution at various concentration and time points was pipetted into each cuvette for measurement. | ||
- | ** The time points include 0, 0.5, 1, 1.5, 2, 2.5, 3.5 | + | ** The time points include 0, 0.5, 1, 1.5, 2, 2.5, and 3.5 hours |
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OD600 of the BL21 liquid culture overnight was 1.063. Surprisingly, this is different than our results from [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.13 Determining E coli concentration with spectrophotometer|5.13 Determining E coli Concentration With Spectrophotometer]]. This implies that we'll have to take several more measurements with overnight liquid culture to determine whether the concentration of bacteria in liquid culture is stable. | OD600 of the BL21 liquid culture overnight was 1.063. Surprisingly, this is different than our results from [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.13 Determining E coli concentration with spectrophotometer|5.13 Determining E coli Concentration With Spectrophotometer]]. This implies that we'll have to take several more measurements with overnight liquid culture to determine whether the concentration of bacteria in liquid culture is stable. | ||
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+ | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
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|0.271 | |0.271 | ||
|0.185 | |0.185 | ||
- | |0. | + | |0.105 |
|- | |- | ||
|1.5 h | |1.5 h | ||
- | |0. | + | |0.326 |
- | |0. | + | |0.223 |
- | |0. | + | |0.132 |
|- | |- | ||
|2 h | |2 h | ||
- | |0. | + | |0.416 |
- | |0. | + | |0.311 |
- | |0. | + | |0.144 |
|- | |- | ||
|2.5 h | |2.5 h | ||
- | |0. | + | |0.485 |
- | |0. | + | |0.426 |
- | |0. | + | |0.179 |
|- | |- | ||
- | |3 h | + | |3.1 h |
- | |0. | + | |0.629 |
- | |0. | + | |0.542 |
- | |0. | + | |0.285 |
|- | |- | ||
|3.5 h | |3.5 h | ||
- | |0. | + | |0.674 |
- | |0. | + | |0.661 |
- | |0. | + | |0.368 |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
|} | |} | ||
- | 2. | + | </center> |
+ | |||
+ | 2. Plots | ||
+ | [[File:BYUSPBacteriaGrowth50.JPG|400px|center|link=]] | ||
- | + | [[File:BYUSPBacteriaGrowth100.JPG|400px|center|link=]] | |
+ | [[File:BYUSPBacteriaGrowth500.JPG|400px|center|link=]] | ||
'''VI) Conclusion''' | '''VI) Conclusion''' | ||
+ | : This experiment indicates that BL21 overnight liquid culture might not have the same concentration of bacteria. We'll need to measure bacterial concentration using spectrophotometer every time during mutagenesis to get an accurate estimate of the number of bacteria we are adding. However, to get approximately OD 0.5, for our dilution step in our mutagenesis protocol, we will use 1:50. | ||
</font> | </font> |
Latest revision as of 15:37, 9 September 2013
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7.27 Bacteria Growth Curve After Dilution
I) Purpose
1. Bacteria preparation
2. Measurements using spectrophotometer
1. Raw data OD600 of the BL21 liquid culture overnight was 1.063. Surprisingly, this is different than our results from 5.13 Determining E coli Concentration With Spectrophotometer. This implies that we'll have to take several more measurements with overnight liquid culture to determine whether the concentration of bacteria in liquid culture is stable.
2. Plots
VI) Conclusion
|