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Revision as of 15:51, 29 July 2014 (view source) SC (Talk | contribs) (→PCR of BBa_K762100) ← Older edit		Revision as of 16:10, 29 July 2014 (view source) Maschi (Talk | contribs) (→Tuesday 29th July) Newer edit →
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	====Electrophoresis====		====Electrophoresis====
-	''by Fabio & Romain''	+	''by Romain''
		+
		+	===E - Salicylate Inducible Suppressing System===
		+	====Digestion====
		+	''by Fabio''
		+
		+	BioBrick '''BBa_J61051''' (Salicylate promoter + NahR)
		+
		+	====Electrophoresis====
		+	''by Fabio''
		+
		+	'''Results:'''
		+
		+	First process to verify the digestion
		+
		+	Second one to separate both parts (only with J61051).
	'''Members there''':		'''Members there''':

---

Revision as of 16:10, 29 July 2014

Contents 1 Tuesday 29th July 1.1 Lab Work 1.1.1 A - The frame 1.1.1.1 PCR 1.1.1.2 Electrophoresis 1.1.2 E - Salicylate Inducible Suppressing System 1.1.2.1 Digestion 1.1.2.2 Electrophoresis 1.1.3 C - Lemon scent 1.1.3.1 PCR of BBa_K762100

Tuesday 29th July

Lab Work

A - The frame

PCR

by Romain

StrainS used: E. coli MG1655Z1 and E. coli MG1655, and oligonucleotides used: FTR-Apra-F and FTR-Apra-R.

Protocol:

• 218µl of H20 MilliQ
• 80µl GoTaq buffer 5X
• 16µl oligonucleotide FTR-Apra-F (10mM)
• 16µl oligonucleotide FTR-Apra-R (10mM)
• 12µl DMSO 10mM
• 32µl MgCl2 (25mM)
• 8µl dNTP (10mM)
• 2µl of GoTaq enzyme
• 2µl of different bacterial cultures in each tubes:

1. Tube 1: MG1655Z1 10I
2. Tube 2: MG1655 100II
3. Tube 3: MG1655 50I
4. Tube 4: MG1655 100I
5. Tube 5: MG1655Z1 10II
6. Tube 6: MG1655 50II
7. Tube 7: MG1655Z1 10I positive control with plasmid

Electrophoresis

by Romain

E - Salicylate Inducible Suppressing System

Digestion

by Fabio

BioBrick BBa_J61051 (Salicylate promoter + NahR)

Electrophoresis

by Fabio

Results:

First process to verify the digestion

Second one to separate both parts (only with J61051).

Members there:

• Instructors and advisors: Alice, Solenne and Sylvie.
• Students: Arnaud, Fabio, Romain, Sean and Terry.

C - Lemon scent

PCR of BBa_K762100

by Sean

Oligonucleotides used: iPS66, iPS67

Oligonucleotides were diluted twice from 100µM to 50µM.

Protocol

Add into a PCR tube the following:

Component	For a total volume of 50μl
H2O	35.5μl
Phusion buffer 5X	10μl
dNTPs	1μl
iPS66	1μl
iPS67	1μl
BBa_K762100	1μl
Phusion DNA polymerase	0.25μl

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