Part:BBa_K1139022:Experience

PlacIq-M13-Plac-GFP on pSB3

3.Spin the overnight culture of the transformed DH5alpha at 9,000g for 1 minute.
4.Pipette the supernatant into a 1.5 mL tube.
5.Dilute it 100 times with water. (=> phage-particle-solution)
6.Melt YT soft agar using a microwave.
7.Dispense 3.5 mL of the melted soft agar to a new round tube, and keep them at 50°C in an incubator.
8.Dispense 400 µL of overnight culture of JM109 to a 1.5 mL tube.
9.Into the 1.5 mL tube, add 100 µL of the phage-particle-solution.
10.Transfer all the content of the 1.5 mL tube to the dispensed soft agar, mix them, and decant all of it on a YT plate.

Result

The result of the plaque forming assay is showed in Fig. 2. M13 phage genes and M13 origin worked together with lacI^q promoter and pSB origin.

Fig. 2-A. The plaques

Fig. 2-B. The negative control

Fig. 2-A.
The plaques were formed using JM109 overnight culture and phage-particle-solution. The phage particles were obtained from the supernatant of the overnight culture of transformed DH5alpha (with pSB3K3- PlacI^q-M13) at 9,000g. Mixture of 3.5 mL YT soft agar, 100 µL of X100 supernatant and 400 µL of overnight culture of JM109 was poured on a YT plate.

Fig. 2-B.
A mixture of only 3.5 mL YT soft agar and 500 µL of overnight culture of JM109 was poured on a YT plate.

For more information, see [http://2013.igem.org/Team:Tokyo_Tech/Experiment/pSB-M13_Plasmid_Assay our work in Tokyo_Tech 2013 wiki].

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UNIQ247fd53112219f81-partinfo-00000003-QINU UNIQ247fd53112219f81-partinfo-00000004-QINU

Bacto tryptone	10 g/L
Yeast Extract	5 g/L
NaCl	10 g/L

Bacto tryptone	8 g/L
Yeast Extract	5 g/L
NaCl	5 g/L
Agarose	15 g/L

Bacto tryptone	8 g/L
Yeast Extract	5 g/L
NaCl	5 g/L
Agarose	6 g/L

Part:BBa_K1139022:Experience

Contents

Materials and Methods

Result

Applications of BBa_K1139022

User Reviews