Revision as of 15:28, 7 August 2015 (view source) JohanB (Talk | contribs) (→‎Wednesday 5th August) ← Older edit		Revision as of 09:38, 10 August 2015 (view source) Coralie (Talk | contribs) (→‎Ligation) Newer edit →
Line 24:		Line 24:
	Biobricks:		Biobricks:
	* BBa_K1707020 : BBa_K1707004 + BBa_K1707019		* BBa_K1707020 : BBa_K1707004 + BBa_K1707019
−
	Mix:		Mix:
−	* 6,5 µL BBa_K1707019	+	** 6,5 µL BBa_K1707019
−	* 9,5 µL BBa_K1707004	+	** 9,5 µL BBa_K1707004
−	* 2 µL Tampon 10x	+	** 2 µL Tampon 10x
−	* 1 µL Ligase	+	** 1 µL Ligase
−	* 1 µL H2O	+	** 1 µL H2O
	* BBa Test : BBa_K1707004 + BBa_K1707013		* BBa Test : BBa_K1707004 + BBa_K1707013
−
	Mix:		Mix:
−	* 5 µL BBa_K1707013	+	** 5 µL BBa_K1707013
−	* 7,5 µL BBa_K1707004	+	** 7,5 µL BBa_K1707004
−	* 1,5 µL Tampon 10x	+	** 1,5 µL Tampon 10x
−	* 1 µL Ligase	+	** 1 µL Ligase
	* BBa_K1707027 : BBa_K1707000 + BBa_E0022		* BBa_K1707027 : BBa_K1707000 + BBa_E0022
−
	Mix:		Mix:
−	* 5 µL BBa_K1707000	+	** 5 µL BBa_K1707000
−	* 9 µL BBa_E0022	+	** 9 µL BBa_E0022
−	* 1,5 µL Tampon 10x	+	** 1,5 µL Tampon 10x
−	* 1 µL Ligase	+	** 1 µL Ligase
−	* 3,5 µL H2O	+	** 3,5 µL H2O
	* BBa_K1707028 : BBa_R0051 + BBa_E0422		* BBa_K1707028 : BBa_R0051 + BBa_E0422
−
	Mix:		Mix:
−	* 6,25 µL BBa_R0051	+	** 6,25 µL BBa_R0051
−	* 5,5 µL BBa_E0422	+	** 5,5 µL BBa_E0422
−	* 1,5 µL Tampon 10x	+	** 1,5 µL Tampon 10x
−	* 1 µL Ligase	+	** 1 µL Ligase
−	* 0,75 µL H2O	+	** 0,75 µL H2O
−		+
	===Rehydratation===		===Rehydratation===

---

Revision as of 09:38, 10 August 2015

Wednesday 5th August

Lab Work

Quantification

by Pauline and Coralie

Biobricks:

• BBa_R0051
• BBa_E0022 (15ng/mL)
• BBa_E0422 (20ng/mL)
• BBa_K1707000 (20ng/mL)
• BBa_K1707004 (15ng/mL)
• BBa_K1707011
• BBa_K1707012
• BBa_K1707013 (20ng/mL)
• BBa_K1707019 (15ng/mL)

Agarose gel, 1% Migration 90V

Ligation

by Pauline

Biobricks:

• BBa_K1707020 : BBa_K1707004 + BBa_K1707019

Mix:

• • 6,5 µL BBa_K1707019
  • 9,5 µL BBa_K1707004
  • 2 µL Tampon 10x
  • 1 µL Ligase
  • 1 µL H2O

• BBa Test : BBa_K1707004 + BBa_K1707013

Mix:

• • 5 µL BBa_K1707013
  • 7,5 µL BBa_K1707004
  • 1,5 µL Tampon 10x
  • 1 µL Ligase

• BBa_K1707027 : BBa_K1707000 + BBa_E0022

Mix:

• • 5 µL BBa_K1707000
  • 9 µL BBa_E0022
  • 1,5 µL Tampon 10x
  • 1 µL Ligase
  • 3,5 µL H2O

• BBa_K1707028 : BBa_R0051 + BBa_E0422

Mix:

• • 6,25 µL BBa_R0051
  • 5,5 µL BBa_E0422
  • 1,5 µL Tampon 10x
  • 1 µL Ligase
  • 0,75 µL H2O

Rehydratation

by Audrey

BBa_R0040

Transformation

by Audrey and Coralie

• BBa_R0040
• BBa Test
• BBa_K1707028
• BBa_K1707027
• BBa_K1707020

Digestion

by Pauline

XbaI + PstI:

• BBa_K1707012
• BBa_K1707011
• BBa_I13600

SpeI + Pst:

• BBa_R0051
• BBa_K1707008

EcoRI + SpeI:

• BBa_K1707004

XbaI + EcoRI:

• BBa_I13602

Soil experiment

by Coralie

Soil

Observation of plates (31/07/2015):

• Soil + H2O on new MCK: lots of colonies are visible
• Soil + LB on new MCK: lots of colonies are visible
• LB on new MCK: nothing
• MCK alone: nothing

We can suppose two different hypothesis:

• The soil is contaminated
• the MCK isn't OK for our experiment: now we add lactose in the preparation. We will try with sorbitol instead of lactose.

We try soil + H2O and soil +LB on:

• MCK + Sorbitol: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol
• MCK + Lactose: Without antibiotics, with Spectinomycine, Tetramycine or Chloramphenicol

Incubation ON, 37°C

Member present:

• Instructors: Claire
• Students: Coralie, Audrey and Pauline

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