2012 Promega catalogue

Life
Science
Catalog
2012
Worldwide Contact List
Section
Contents
Table of
Contents
Cell Signaling
104
pSP73 Vector
Product Size Cat.# Price ($)
pSP73 Vector 20 µg P2221 143.00
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The pSP73 Vector offers a wide range of restriction sites,
providing greater versatility in cloning and transcription of RNA in vitro. The
pSP73 Vector contains the SP6 and T7 RNA polymerase promoters and a
unique multiple cloning region, which includes restriction sites for BglII, EcoRV,
ClaI, EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, AccI, PstI, SphI, HindIII, PvuII
and XhoI. The pSP72 and pSP73 Vectors are essentially identical except for
the orientation of the multiple cloning region.
Features:
• Versatile: This vector can be used for standard cloning and in vitro
transcription from SP6 and T7 RNA polymerase promoters flanking the
multiple cloning region.
• Convenient: Multiple cloning site provides a selection of restriction sites
for cloning.
Storage Conditions: Store vector at –20°C.
Protocol Part#
pSP73 Vector Technical Bulletin TB041
BglI
1330
XmnI 1809
ScaI 1690
PvuI 1580
FspI
1432
Amp r
pSP73
Vector
(2,464bp)
ori
SspI 2014
AatII 2132
NdeI
2381
HpaI
138
SP6
BglII
EcoRV
ClaI
EcoRI
SacI
KpnI
SmaI
BamHI
XbaI
SalI
AccI
PstI
SphI
HindIII
PvuII
XhoI
T7
➞➞
1 start
6
14
19
24
34
40
42
45
51
57
58
67
73
75
83
87
103
0300VA05_2A
Bacterial Strains and Competent Cells
Product
Bacterial Strain ES1301 mutS, Glycerol Stock
Size Cat.# Price ($)
(noncompetent)
Bacterial Strain BMH 71-18 mutS, Glycerol Stock
200 µl Q6131 69.00
(noncompetent) 500 µl Q6321 69.00
Bacterial Strain JM109, Glycerol Stock 500 µl P9751 67.00
Bacterial Strain JM109(DE3), Glycerol Stock 500 µl P9801 67.00
Bacterial Strain LE392, Glycerol Stock 500 µl K9981 66.00
Bacterial Strain NM522, Glycerol Stock 500 µl P2301 67.00
Single Step (KRX) Competent Cells 20 × 50 µl L3002 330.00
L-Rhamnose Monohydrate 10 g L5701 70.00
50 g L5702 272.00
JM109 Competent Cells, >107cfu/µg 1 ml L1001 145.00
JM109 Competent Cells, >108cfu/µg 1 ml L2001 195.00
HB101 Competent Cells, >108cfu/µg 1 ml L2011 170.00
BL21(DE3)pLysS Competent Cells, >106cfu/µg 1 ml L1191 238.00
A.
Normalized luminescence (RLU/OD)
B.
Induction Ratio
10,000
9,000
8,000
7,000
6,000
5,000
4,000
3,000
2,000
1,000
0
10,000
1,000
100
10
1
Normalized luminescence (RLU/OD)
14
140
120
100
80
60
40
20
0
BL21(DE3) BL21(DE3)
pLysS
For complete and up-to-date product information visit: www.promega.com/catalog
8
BL21(DE3) BL21(DE3)
pLysS
Pre-Induction Expression
Roset t a 2
pLysS
BL21(DE3)
pLysS
BL21(DE3)
43
Rosetta 2
pLysS
KRX
Rosetta 2
pLysS
1,700
KRX
KRX
Pre-induction and post-induction expression levels of firefly
luciferase. Cells were transformed with the pF1K T7 Flexi ® Vector
containing the firefly luciferase gene. Cultures were grown at 37°C to
an optical density (O.D. 600) of 0.8–1.0 and then moved to a 25°C incubator
shaker. When cultures reached an O.D. 600 of 1.0–1.5, protein expression
was induced using either 0.1% rhamnose or 1mM IPTG and grown overnight
at 25°C. Samples for luciferase assays were removed prior to and after
induction. Panel A. Firefly luciferase expression level was determined using
the Bright-Glo Luciferase Assay Reagent. Pre- and post-induction firefly
luciferase expression levels were normalized to cell number (n = 3). Panel B.
Induction ratios were calculated by dividing the post-induction luminescence
values by the pre-induction values.
6032MA