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Team:Hawaii/Notebook/2008-06- 3
From 2008.igem.org
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== Things we did today == | == Things we did today == | ||
| + | '''Grace & Norman''' | ||
* Thaw test for PCC6803 and DH5-α | * Thaw test for PCC6803 and DH5-α | ||
* Made LB and LB + amp media | * Made LB and LB + amp media | ||
| + | :* amp conc. = 100 μg/mL | ||
* Made 1000X amp stock solution, stored in 4C freezer | * Made 1000X amp stock solution, stored in 4C freezer | ||
| + | :* 1000X = 100 mg/mL | ||
* Tested competency of DH5-α [[Team:Hawaii/Test_Competent_E._Coli | (see experiment)]] | * Tested competency of DH5-α [[Team:Hawaii/Test_Competent_E._Coli | (see experiment)]] | ||
* Counted colonies to determine number of cells grown to OD600 = 0.3 | * Counted colonies to determine number of cells grown to OD600 = 0.3 | ||
| Line 13: | Line 16: | ||
{| border="1" | {| border="1" | ||
|+ '''Cell Count''' | |+ '''Cell Count''' | ||
| - | !width=" | + | !width="225"|Dilution |
| - | !width=" | + | !width="225"|Colony Forming Units on Plate |
| - | !width=" | + | !width="225"|Number of Cells per mL |
|- | |- | ||
! 1 | ! 1 | ||
| - | | Too many to count || n/a | + | | align="center"|Too many to count || align="center"|n/a |
|- | |- | ||
! 1/10 | ! 1/10 | ||
| - | |Too many to count | + | |align="center"|Too many to count |
| - | |n/a | + | |align="center"|n/a |
|- | |- | ||
! 1/100 | ! 1/100 | ||
| - | | Too many to count|| n/a | + | | align="center"|Too many to count|| align="center"|n/a |
|- | |- | ||
! 1/1000 | ! 1/1000 | ||
| - | | 190|| 19,000,000 | + | | align="center"|190|| align="center"|19,000,000 |
|- | |- | ||
! 1/10,000 | ! 1/10,000 | ||
| - | | 40|| 4,000,000 | + | | align="center"|40|| align="center"|4,000,000 |
|- | |- | ||
! 1/100,000 | ! 1/100,000 | ||
| - | | 23|| 23,000,000 | + | | align="center"|23|| align="center"|23,000,000 |
|- | |- | ||
! 1/1,000,000 | ! 1/1,000,000 | ||
| - | | 6|| | + | | align="center"|6|| align="center"|600,000 |
|} | |} | ||
| - | + | [[Image:DSC00696.JPG|left|thumb|250px|DH5-α dilutions plated on LB]] | |
| - | [[Image:DSC00696.JPG| | + | <br> |
== Discussion == | == Discussion == | ||
We have lots of cells (yay!). Cryofreezing will result in the loss cell population, but we will very likely still have viable cells after thawing. | We have lots of cells (yay!). Cryofreezing will result in the loss cell population, but we will very likely still have viable cells after thawing. | ||
| + | |||
| + | == Quote of the Day == | ||
| + | |||
| + | "Wow, competency takes a long time." - NW | ||
{{Team:Hawaii/Footer}} | {{Team:Hawaii/Footer}} | ||
Latest revision as of 02:34, 5 June 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Grace & Norman
- Thaw test for PCC6803 and DH5-α
- Made LB and LB + amp media
- amp conc. = 100 μg/mL
- Made 1000X amp stock solution, stored in 4C freezer
- 1000X = 100 mg/mL
- Tested competency of DH5-α (see experiment)
- Counted colonies to determine number of cells grown to OD600 = 0.3
- Cells were cryoperserved, OD600 = 0.0748
- 10 μl were plated on each plate
| Dilution | Colony Forming Units on Plate | Number of Cells per mL |
|---|---|---|
| 1 | Too many to count | n/a |
| 1/10 | Too many to count | n/a |
| 1/100 | Too many to count | n/a |
| 1/1000 | 190 | 19,000,000 |
| 1/10,000 | 40 | 4,000,000 |
| 1/100,000 | 23 | 23,000,000 |
| 1/1,000,000 | 6 | 600,000 |
Discussion
We have lots of cells (yay!). Cryofreezing will result in the loss cell population, but we will very likely still have viable cells after thawing.
Quote of the Day
"Wow, competency takes a long time." - NW
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
