Team:Paris/August 1

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Latest revision as of 16:35, 23 August 2008

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MiniPreps

Conditions

Use of Quiagen's protocol on all the clones cultivated on the 31th.
Preparation of 50µl of minipreps in water.

List of Minipreps

Name	Biobricks	Description
MP124	Strong rbs - YFP D129 (BV) - D118 (BI)
MP125	Strong rbs - mRFP D129 (BV) - D122 (BI)
MP126	Strong rbs - lasR activator D129 (BV) - D114 (BI)
MP127	Strong promoter - ECFP D123 (BV) - D130 (BI)
MP128	Strong promoter - gfp Tripart D123 (BV) - D131 (BI)
MP129	Strongest promoter - ECFP D103 (BV) - D131 (BI)
MP130	Strong promoter - gfp Tripart D103 (BV) - D130 (BI)
MP131	Medium promoter - gfp Tripart D124 (BV) - D131 (BI)
MP132	Weak promoter - ECFP D104 (BV) - D130 (BI)
MP133	Weak promoter - gfp D104 (BV) - D131 (BI)
MP134	pLas - ECFP D105 (BV) - D130 (BI)
MP135	pLas - gfp Tripart D105 (BV) - D131 (BI)
MP136	yfp - Double Terminator D125 (FV) - D117 (FI)
MP137	rfp - Double Terminator D125 (FV) - D121 (FI)
MP138	lasR activator + LVA - Double Terminator D125 (FV) - D113 (FI)
MP139	Strong promoter - gfp tripart D106 (BV) - D131 (BI)
MP140	Strongest RBS - rfp D102 (BV) - D122 (BI)
MP141	Strongest RBS - yfp D102 (BV) - D118 (BI)

Glycerol Stock

Protocol

1mL glycerol
1mL culture in LB

List of Strain stocked

Strain	Ligation	Biobricks	Description
S123	L102	Strong rbs - YFP D129 (BV) - D118 (BI)
S124	L103	Strong rbs - mRFP D129 (BV) - D122 (BI)
S125	L104	Strong rbs - lasR activator D129 (BV) - D114 (BI)
S126	L105	Strong promoter - ECFP D123 (BV) - D130 (BI)
S127	L106	Strong promoter - gfp Tripart D123 (BV) - D131 (BI)
S128	L107	Strongest promoter - gfp Tripart D103 (BV) - D131 (BI)
S129	L108	Weak promoter - ECFP D103 (BV) - D130 (BI)
S130	L110	Medium promoter - gfp D124 (BV) - D131 (BI)
S131	L111	Weak promoter - ECFP D104 (BV) - D130 (BI)
S132	L112	Weak promoter - gfp Tripart D104 (BV) - D131 (BI)
S133	L115	pLas - ECFP D105 (BV) - D130 (BI)
S134	L116	pLas - gfp Tripart D105 (BV) - D131 (BI)
S135	L117	yfp - Double Terminator D125 (FV) - D117 (FI)
S136	L118	rfp - Double Terminator D125 (FV) - D117 (FI)
S137	L119	LasR activator + LVA - Double terminator D125 (FV) - D113 (FI)
S138	L121	Strong promoter - gfp Tripart D106 (BV) - D130 (BI)
S139	L124	Strongest rbs - mRFP D102 (BV) - D122 (BI)
S140	L125	Strongest rbs - yfp D102 (BV) - D118 (BI)

Transformations

Protocol

use of TOP10 Chemically competent cells

Defroze competent cells on ice during 5'
Add 5µl of DNA Ligation in 50µL of competent bacterias (or 1µL for the positive control puc19)
Incubate 30' on ice
Heat-shock the cells during 30" at 42°C without shaking
Put 2' on ice
Add 250µL of pre-warmed SOC medium (4°C)
Incubate 1h at 37°C under shaking (225rpm)
Spin at 5.000rpm during 30"
Remove 150µL of supernatant
Resuspent the pellet in the 150µL left
Spread on adequated plates
Incubate O/N at 37°C

List of transformation

Name	Description	Biobrick
L100	RBS-TetR-ECFP
L101	RBS-tetR-GFP tripart
L113	pBAD-ECFP
L114	pBAD-GFP tripart
L120	ptet-ECFP
L122	RBS-lasI-ECFP
L123	RBS-lasI-GFP-ter
L126	RBS-lasR-LVA

@@ Line 10: / Line 10: @@
+=== List of Minipreps ===
 {| border="1"
 |align="center"|'''Name'''
-|align="center"|'''Description'''
 |align="center"|'''Biobricks'''
+|align="center"|'''Description'''
 |-
-|align="center"|MP L102
+|align="center"|MP124
+|align="center"|Strong rbs - YFP<br>D129 (BV) -  D118 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-|align="center"|
-Strong rbs - YFP
-<br>D129 (BV) -  D118 (BI)
 |-
-|align="center"|MP L103
+|align="center"|MP125
+|align="center"|Strong rbs - mRFP<br>D129 (BV) -  D122 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-|align="center"|
-Strong rbs - GFP
-<br>D129 (BV) -  D122 (BI)
 |-
-|align="center"|MP L104
+|align="center"|MP126
+|align="center"|Strong rbs - lasR activator<br>D129 (BV) -  D114 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
-|align="center"|
-Strong rbs - lasR activator
-<br>D129 (BV) -  D114 (BI)
 |-
-|align="center"|MP L105
+|align="center"|MP127
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
-|align="center"|
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Strong promoter - ECFP
-<br>D123 (BV) - D130 (BI)
 |-
-|align="center"|MP L106
+|align="center"|MP128
+|align="center"|Strong promoter - gfp Tripart<br>D123 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-|align="center"|
-Strong promoter - gfp Tripart
-<br>D123 (BV) - D131 (BI)
 |-
-|align="center"|MP L107
+|align="center"|MP129
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|Strongest promoter - ECFP<br>D103 (BV) - D131 (BI)
-|align="center"|
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Strongest promoter - ECFP
-<br>D103 (BV) - D131 (BI)
 |-
-|align="center"|MP L108
+|align="center"|MP130
+|align="center"|Strong promoter -  gfp Tripart<br>D103 (BV) - D130 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-|align="center"|
-Strong promoter -  gfp Tripart
-<br>D103 (BV) - D130 (BI)
 |-
-|align="center"|MP L110
+|align="center"|MP131
+|align="center"|Medium promoter - gfp Tripart<br>D124 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-|align="center"|
-Medium promoter - gfp Tripart
-<br>D124 (BV) - D131 (BI)
 |-
-|align="center"|MP L111
+|align="center"|MP132
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)
-|align="center"|
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Weak promoter - ECFP
-<br>D104 (BV) - D130 (BI)
 |-
-|align="center"|MP L112
+|align="center"|MP133
+|align="center"|Weak promoter - gfp<br>D104 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-|align="center"|Weak promoter - gfp
-<br>D104 (BV) - D131 (BI)
 |-
-|align="center"|MP L115
+|align="center"|MP134
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|pLas - ECFP<br>D105 (BV) - D130 (BI)
-|align="center"|
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-pLas - ECFP
-<br>D105 (BV) - D130 (BI)
 |-
-|align="center"|MP L116
+|align="center"|MP135
+|align="center"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-|align="center"|
-pLas - gfp Tripart
-<br>D105 (BV) - D131 (BI)
 |-
-|align="center"|MP L117
+|align="center"|MP136
-|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
+|align="center"|yfp - Double Terminator<br>D125 (FV) - D117 (FI)
-|align="center"|
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-Double Terminator - yfp
-<br>D125 (FV) - D117 (FI)
 |-
-|align="center"|MP L118
+|align="center"|MP137
-|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
+|align="center"|rfp - Double Terminator<br>D125 (FV) - D121 (FI)
-|align="center"|
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-Double Terminator - rfp
-<br>D125 (FV) - D121 (FI)
 |-
-|align="center"|MP L119
+|align="center"|MP138
-|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Icon_coding.png]]
+|align="center"|lasR activator + LVA - Double Terminator<br>D125 (FV) - D113 (FI)
-|align="center"|
+|align="center"|[[Image:Icon_coding.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-Double Terminator - lasR activator + LVA
-<br>D125 (FV) - D113 (FI)
 |-
-|align="center"|MP L121
+|align="center"|MP139
+|align="center"|Strong promoter - gfp tripart<br>D106 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-|align="center"|
-Strong promoter - gfp tripart
-<br>D106 (BV) - D130 (BI)
 |-
-|align="center"|MP L124
+|align="center"|MP140
+|align="center"|Strongest RBS - rfp<br>D102 (BV) - D122 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-|align="center"|
-Strongest RBS - rfp
-<br>D102 (BV) - D122 (BI)
 |-
-|align="center"|MP L125
+|align="center"|MP141
+|align="center"|Strongest RBS - yfp<br>D102 (BV) - D118 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-|align="center"|
-Strongest RBS - yfp
-<br>D102 (BV) - D118 (BI)
 |}
+== Glycerol Stock ==
-== Stock Glycerol ==
 === Protocol ===
@@ Line 132: / Line 96: @@
 * 1mL culture in LB
+=== List of Strain stocked ===
 {|border="1"
-|Strain
+|align="center"|'''Strain'''
-|Ligation
+|align="center"|'''Ligation'''
-|Description
+|align="center"|'''Biobricks'''
+|align="center"|'''Description'''
 |-
-|S123
+|align="center"|S123
+|align="center"|L102
+|align="center"|Strong rbs - YFP<br>D129 (BV) -  D118 (BI)
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
+|-
+|align="center"|S124
+|align="center"|L103
+|align="center"|Strong rbs - mRFP<br>D129 (BV) - D122 (BI)
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
+|-
+|align="center"|S125
 |align="center"|L104
+|align="center"|Strong rbs - lasR activator<br>D129 (BV) - D114 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
-Strong rbs - lasR activator
+|-
-<br>D129 (BV) -  D114 (BI)
+|align="center"|S126
-|-
-|S124
 |align="center"|L105
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|Strong promoter - ECFP<br>D123 (BV) - D130 (BI)
-Strong promoter - ECFP
-<br>D123 (BV) - D130 (BI)
-|-
-|S125
-|align="center"|L106
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Strong promoter - gfp Tripart
-<br>D123 (BV) - D131 (BI)
-|-
-|S126
-|align="center"|L107
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-Strongest promoter - ECFP
-<br>D103 (BV) - D131 (BI)
 |-
-|S127
+|align="center"|S127
-|align="center"|L108
+|align="center"|L106
+|align="center"|Strong promoter -  gfp Tripart<br>D123 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Strong promoter -  gfp Tripart
-<br>D103 (BV) - D130 (BI)
 |-
-|S128
+|align="center"|S128
-|align="center"|L110
+|align="center"|L107
+|align="center"|Strongest promoter - gfp Tripart<br>D103 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Medium promoter - gfp Tripart
-<br>D124 (BV) - D131 (BI)
 |-
-|S129
+|align="center"|S129
-|align="center"|L111
+|align="center"|L108
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|Weak promoter - ECFP<br>D103 (BV) - D130 (BI)
-Weak promoter - ECFP
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-<br>D104 (BV) - D130 (BI)
 |-
-|S130
+|align="center"|S130
-|align="center"|L112
+|align="center"|L110
+|align="center"|Medium promoter - gfp<br>D124 (BV) - D131 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
-Weak promoter - gfp
+|-
-<br>D104 (BV) - D131 (BI)
+|align="center"|S131
+|align="center"|L111
+|align="center"|Weak promoter - ECFP<br>D104 (BV) - D130 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
-|- style="background: #dddddd;"
+|align="center"|S132
-| style="background: #D4E2EF;" |S131
+|align="center"|L112
+|align="center"|Weak promoter - gfp Tripart<br>D104 (BV) - D131 (BI)
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
+|-
+|align="center"|S133
 |align="center"|L115
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
+|align="center"|pLas - ECFP <br>D105 (BV) - D130 (BI)
-pLas - ECFP
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-<br>D105 (BV) - D130 (BI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S134
-| style="background: #D4E2EF;" |S132
 |align="center"|L116
-|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
+|align="center"|pLas - gfp Tripart<br>D105 (BV) - D131 (BI)
-pLas - gfp Tripart
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-<br>D105 (BV) - D131 (BI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S135
-| style="background: #D4E2EF;" |S133
 |align="center"|L117
-|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
+|align="center"|yfp - Double Terminator<br>D125 (FV) - D117 (FI)
-Double Terminator - yfp
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-<br>D125 (FV) - D117 (FI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S136
-| style="background: #D4E2EF;" |S134
 |align="center"|L118
-|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_reporter.png]]
+|align="center"|rfp - Double Terminator<br>D125 (FV) - D117 (FI)
-Double Terminator - rfp
+|align="center"|[[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-<br>D125 (FV) - D121 (FI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S137
-| style="background: #D4E2EF;" |S135
 |align="center"|L119
-|align="center"|[[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]][[Image:Icon_coding.png]]
+|align="center"|LasR activator + LVA - Double terminator<br>D125 (FV) - D113 (FI)
-Double Terminator - lasR activator + LVA
+|align="center"|[[Image:Icon_coding.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
-<br>D125 (FV) - D113 (FI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S138
-| style="background: #D4E2EF;" |S136
 |align="center"|L121
+|align="center"|Strong promoter - gfp Tripart<br>D106 (BV) - D130 (BI)
 |align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
-Strong promoter - gfp tripart
-<br>D106 (BV) - D130 (BI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S139
-| style="background: #D4E2EF;" |S137
 |align="center"|L124
+|align="center"|Strongest rbs - mRFP<br>D102 (BV) - D122 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-Strongest RBS - rfp
-<br>D102 (BV) - D122 (BI)
 |-
-|- style="background: #dddddd;"
+|align="center"|S140
-| style="background: #D4E2EF;" |S138
 |align="center"|L125
+|align="center"|Strongest rbs - yfp<br>D102 (BV) - D118 (BI)
 |align="center"|[[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]]
-Strongest RBS - yfp
-<br>D102 (BV) - D118 (BI)
 |}
 == Transformations ==
 === Protocol ===
+''use of TOP10 Chemically competent cells''
-use of TOP10 Chemically competent cells
 * Defroze competent cells on ice during 5'
@@ Line 266: / Line 217: @@
 === List of transformation ===
 {| border="1"
 |align="center"|'''Name'''
+|align="center"|'''Description'''
 |align="center"|'''Biobrick'''
-|align="center"|'''Description'''
 |-
 |align="center"|L100
-|align="center"|
 |align="center"| RBS-TetR-ECFP
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|L101
-|align="center"|
 |align="center"|RBS-tetR-GFP tripart
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|L113
-|align="center"|
 |align="center"|pBAD-ECFP
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|L114
-|align="center"|
+|align="center"|pBAD-GFP tripart
-|align="center"|pBAD-GFP
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_reporter.png]]
 |-
 |align="center"|L120
-|align="center"|
+|align="center"|ptet-ECFP
-|align="center"|tetR-ECFP
+|align="center"|[[Image:Part_icon_regulatory.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|L122
-|align="center"|
 |align="center"|RBS-lasI-ECFP
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]][[Image:Part_icon_rbs.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|L123
-|align="center"|
 |align="center"|RBS-lasI-GFP-ter
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]]
 |-
 |align="center"|L126
-|align="center"|
 |align="center"|RBS-lasR-LVA
+|align="center"|[[Image:Part_icon_rbs.png]][[Image:Icon_coding.png]]
 |}

Team:Paris/August 1

From 2008.igem.org

Latest revision as of 16:35, 23 August 2008

Contents

MiniPreps

Conditions

List of Minipreps

Glycerol Stock

Protocol

List of Strain stocked

Transformations

Protocol

List of transformation