"
Team:Kyoto/Notebook
From 2010.igem.org
(Difference between revisions)
| Line 3: | Line 3: | ||
== Notebook == | == Notebook == | ||
| + | ===001: Preparing=== | ||
| + | * By: Wataru, Tomo, Yuki, Kazuya, Ken, Makoto | ||
| + | * Date: 07/20 | ||
| + | * Category: Antibiotic, Parts, Transformation | ||
| + | * Protocol: [[Team:Kyoto/Protocol#Use_BioBrick_Parts|Use BioBrick Parts]] | ||
| + | |||
| + | ====1. Solubilization fo antibiotics.==== | ||
| + | # 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). | ||
| + | # Dispense 1.1ml to 1.5ml tubes. | ||
| + | # Keep in -20℃ Freezer. | ||
| + | |||
| + | ====2. Make LB plate.==== | ||
| + | 9 plates for Ampicillin+ and Kanamycin+. | ||
| + | |||
| + | ====3. Transformation of iGEM Parts.==== | ||
| + | {| class="note" | ||
| + | |Name||Well<sup>*1</sup>||Sample (µl)||Competent Cells (µl)||Total (µl)||Plate||Incubation||Result| | ||
| + | |- | ||
| + | |<partinfo>J23100</partinfo>||1-18-C|1||20||21||rowspan="7"|LB (Ampicillin+)||rowspan="8"|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. | ||
| + | |||
| + | ====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". | ||
---- | ---- | ||
Revision as of 14:46, 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
1. Solubilization fo antibiotics.
- 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).
- Dispense 1.1ml to 1.5ml tubes.
- Keep in -20℃ Freezer.
2. Make LB plate.
9 plates for Ampicillin+ and Kanamycin+.
3. Transformation of iGEM Parts.
| Name | Well*1 | Sample (µl) | Competent Cells (µl) | Total (µl) | Plate | Incubation | |
| <partinfo>J23100</partinfo> | 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.
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".
