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Team:Kyoto/Notebook
From 2010.igem.org
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* Protocol: [[Team:Kyoto/Protocol#Use_BioBrick_Parts|Use BioBrick Parts]] | * Protocol: [[Team:Kyoto/Protocol#Use_BioBrick_Parts|Use BioBrick Parts]] | ||
| - | ==== | + | ====Solubilization fo antibiotics==== |
| - | * 1.Make two mixtures as the following. | + | * 1. Make two mixtures as the following. |
** 1g Ampicillin and 20ml MilliQ (Final concentration 50mg/ml). | ** 1g Ampicillin and 20ml MilliQ (Final concentration 50mg/ml). | ||
| - | ** 0 | + | ** 0.5g Ampicillin and 10ml MilliQ (Final concentration 50mg/ml). |
| - | * 2.Dispense 1.1ml to 1.5ml tubes. | + | * 2. Dispense 1.1ml to 1.5ml tubes. |
| - | * 3.Keep in -20℃ Freezer. | + | * 3. Keep in -20℃ Freezer. |
====2. Make LB plate.==== | ====2. Make LB plate.==== | ||
Revision as of 16:01, 24 August 2010
LastModified: 2010-8-24
Contents |
Index
Notebook
001: Preparing
- By: Wataru, Tomo, Yuki, Kazuya, Ken, Makoto
- Date: 07/20
- Category: Antibiotic, Parts, Transformation
- Protocol: Use BioBrick Parts
Solubilization fo antibiotics
- 1. Make two mixtures as the following.
- 1g Ampicillin and 20ml MilliQ (Final concentration 50mg/ml).
- 0.5g Ampicillin and 10ml MilliQ (Final concentration 50mg/ml).
- 2. Dispense 1.1ml to 1.5ml tubes.
- 3. Keep in -20℃ Freezer.
2. Make LB plate.
Make 9 plates for LB (Ampicillin+) and LB (Kanamycin+).
3. Transformation of iGEM Parts.
| Name | Well*1 | Sample (µl) | Competent Cells (µl) | Total (µl) | Plate | Incubation | Result |
| <partinfo>J23100</partinfo> | 1-18-C | 1 | 20 | 21 | LB (Ampicillin+) | At 37℃ 7/20 20:50 - 7/21 17:00 | ○ |
| <partinfo>J23105</partinfo> | 1-18-M | 1 | 20 | 21 | ○ | ||
| <partinfo>J23116</partinfo> | 1-20-M | 1 | 20 | 21 | ○ | ||
| <partinfo>R0011</partinfo> | 1-6-G | 1 | 20 | 21 | ○ | ||
| <partinfo>E0840</partinfo> | 1-12-O | 1 | 20 | 21 | ○ | ||
| <partinfo>J06702</partinfo> | 2-8-E | 1 | 20 | 21 | ○ | ||
| <partinfo>pSB4K5</partinfo> | 1-5-G | 1 | 20 | 21 | × | ||
| <partinfo>B0015</partinfo> | 1-23-L | 1 | 20 | 21 | LB (Kanamycin+) | × |
- *1 "1-18-C" means well 18C in [http://partsregistry.org/Help:Spring_2010_DNA_distribution Spring 2010 DNA Distribution Kit] Plate 1.
4. Discussion
A vector of "pSB4K5" is Kanamycin-resistance, however, we plated it to LB plate (Ampicillin+).
And We didn't pre-culture "B0015" despite its vector is Kanamycin-resistance.
So, it was predicted that we will fail the transformation of "pSB4K5" and "B0015".
