Team:Kyoto/Notebook

From 2010.igem.org

(Difference between revisions)
(Discussion)
(1. Solubilization fo antibiotics.)
Line 10: Line 10:
====1. Solubilization fo antibiotics.====
====1. Solubilization fo antibiotics.====
-
# 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).
-
#* 0,5g Ampicillin and 10ml MilliQ (Final concentration 50mg/ml).
+
** 0,5g Ampicillin and 10ml MilliQ (Final concentration 50mg/ml).
-
# Dispense 1.1ml to 1.5ml tubes.
+
* 2.Dispense 1.1ml to 1.5ml tubes.
-
# Keep in -20℃ Freezer.
+
* 3.Keep in -20℃ Freezer.
====2. Make LB plate.====
====2. Make LB plate.====

Revision as of 15:58, 24 August 2010

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.

  • 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.

9 plates for Ampicillin+ and Kanamycin+.

3. Transformation of iGEM Parts.

NameWell*1Sample (µl)Competent Cells (µl)Total (µl)PlateIncubationResult
<partinfo>J23100</partinfo>1-18-C12021LB (Ampicillin+)At 37℃ 7/20 20:50 - 7/21 17:00
<partinfo>J23105</partinfo>1-18-M12021
<partinfo>J23116</partinfo>1-20-M12021
<partinfo>R0011</partinfo>1-6-G12021
<partinfo>E0840</partinfo>1-12-O12021
<partinfo>J06702</partinfo>2-8-E12021
<partinfo>pSB4K5</partinfo>1-5-G12021×
<partinfo>B0015</partinfo>1-23-L12021LB (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".