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Virginia Commonwealth/4 August 2009
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==Tuesday 4 August 2009== | ==Tuesday 4 August 2009== | ||
===Results=== | ===Results=== | ||
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** Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink | ** Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink | ||
** Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP | ** Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP | ||
| - | [[User:Trentay|Trentay]] 22: | + | * There is a possibility the pSB4C5 backbone may be a bad part |
| + | ** previous years it has not functioned properly | ||
| + | ** Gel electrophoresis indicates DNA Gel Purification may be necessary | ||
| + | * Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing | ||
| + | ** They will be aliquoted into smaller amounts | ||
| + | * Gel Electrophoresis Results | ||
| + | [[Image:2009-08-04 DigestI1352andpSB4C5.jpg|350px|thumb|none| From Left to Right: 100bp Ladder, Bioline Hyperladder, I1352(Digest), I1352(Miniprep), pSB4C5(Digest), pSB4C5(Miniprep)]] | ||
| + | |||
| + | [[User:Trentay|Trentay]] 22:25, 4 August 2009 (UTC) | ||
---- | ---- | ||
| + | |||
===Tasks=== | ===Tasks=== | ||
''Maria and Afton'' | ''Maria and Afton'' | ||
| Line 15: | Line 62: | ||
====Wetlab==== | ====Wetlab==== | ||
* New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts | * New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts | ||
| - | + | {|border="1" | |
| - | + | |+ | |
| - | + | |- | |
| - | + | ! Name !! Amount/Tube !! # Tubes !! Extra in Stock !! | |
| - | + | |- | |
| - | + | ! XbaI | |
| - | + | | 10 µL || 15 || No || | |
| + | |- | ||
| + | ! SpeI | ||
| + | | 5 µL || 10 || No || | ||
| + | |- | ||
| + | ! PstI | ||
| + | | 10 µL || 25 || Yes || | ||
| + | |- | ||
| + | ! EcoRI-HF | ||
| + | | 10 µL || 25 || Yes || | ||
| + | |- | ||
| + | ! BSA | ||
| + | | 5 µL || 8 || Yes || | ||
| + | |- | ||
| + | ! T4 Ligase Buffer | ||
| + | | 10 µL || 25 || Yes || | ||
| + | |- | ||
| + | ! T4 Ligase | ||
| + | | 10 µL || 8 || No || | ||
| + | |} | ||
| + | * To show how many times a tube has been thawed and refrozen, a hash mark will be placed on the side of the tube, each time it is thawed as a reference for others to see | ||
| + | [[User:Trentay|Trentay]] 15:24, 5 August 2009 (UTC) | ||
''Maria and Afton'' | ''Maria and Afton'' | ||
| - | * | + | * Gel Electrophoresis was run |
| + | ** pSB4C5 and I1352 digested and undigested (right after Miniprep) were run on a gel and results showed excessive bands in the undigested DNA. DNA Gel Purification will be done on future Miniprepped DNA | ||
| + | * pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer. | ||
| + | * Transformation was done using DB 3.1 | ||
| + | {|border="1" | ||
| + | |+ | ||
| + | |- | ||
| + | ! Name !! Plate !! Growth Observation !! | ||
| + | |- | ||
| + | ! pSB1C3 | ||
| + | | CM|| n/a || | ||
| + | |- | ||
| + | ! (+) Control | ||
| + | | LB || n/a || | ||
| + | |- | ||
| + | ! (+) Control zapped | ||
| + | | LB || n/a || | ||
| + | |- | ||
| + | ! (-) Control | ||
| + | | CM || n/a || | ||
| + | |} | ||
| + | [[User:Trentay|Trentay]] 22:33, 4 August 2009 (UTC) | ||
Latest revision as of 18:29, 13 August 2009
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Contents |
Tuesday 4 August 2009
Results
Maria and Afton
- Overnight culture of I1352 plated on AMP:
- Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink
- Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP
- There is a possibility the pSB4C5 backbone may be a bad part
- previous years it has not functioned properly
- Gel electrophoresis indicates DNA Gel Purification may be necessary
- Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing
- They will be aliquoted into smaller amounts
- Gel Electrophoresis Results
Trentay 22:25, 4 August 2009 (UTC)
Tasks
Maria and Afton
- Run a Gel Electrophoresis
- Transform and grow up pSB1C3 with the death gene P1010
Trentay 22:11, 4 August 2009 (UTC)
Wetlab
- New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts
| Name | Amount/Tube | # Tubes | Extra in Stock | |
|---|---|---|---|---|
| XbaI | 10 µL | 15 | No | |
| SpeI | 5 µL | 10 | No | |
| PstI | 10 µL | 25 | Yes | |
| EcoRI-HF | 10 µL | 25 | Yes | |
| BSA | 5 µL | 8 | Yes | |
| T4 Ligase Buffer | 10 µL | 25 | Yes | |
| T4 Ligase | 10 µL | 8 | No |
- To show how many times a tube has been thawed and refrozen, a hash mark will be placed on the side of the tube, each time it is thawed as a reference for others to see
Trentay 15:24, 5 August 2009 (UTC)
Maria and Afton
- Gel Electrophoresis was run
- pSB4C5 and I1352 digested and undigested (right after Miniprep) were run on a gel and results showed excessive bands in the undigested DNA. DNA Gel Purification will be done on future Miniprepped DNA
- pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
- Transformation was done using DB 3.1
| Name | Plate | Growth Observation | |
|---|---|---|---|
| pSB1C3 | CM | n/a | |
| (+) Control | LB | n/a | |
| (+) Control zapped | LB | n/a | |
| (-) Control | CM | n/a |
Trentay 22:33, 4 August 2009 (UTC)
