2013 Promega catalogue

Cell Signaling
Subcloning Tools and Vectors
Subcloning Tools Bundle
Product Size Cat.# Price ($)
Subcloning Tools Bundle 1 each M1060 495
For Research Use Only. Not for Use in Diagnostic Procedures. Product may not be available in
all countries. Please contact your local representative for more information.
Description: Speed your subcloning with these easy-to-use tools. Purchase
the Subcloning Tools Bundle, and get LigaFast Rapid DNA Ligation System,
TSAP Thermosensitive Alkaline Phosphatase, BenchTop 100bp DNA Ladder,
Wizard ® SV Gel and PCR Clean-Up System and PureYield Plasmid Miniprep
System for one low price. It’s like getting the PureYield Plasmid Miniprep
System for free.
Features:
• LigaFast Rapid DNA Ligation System: Rapid room temperature ligations
of vectors and inserts in as little as 5 minutes. Transform competent
bacteria immediately following the reaction.
• TSAP Thermosensitive Alkaline Phosphatase: Use rapid protocol (included)
to digest and dephosphorylate at the same time or use in standard
application. Heat-deactivate the enzyme after the reaction in 15 minutes.
Active in common restriction enzyme buffers with no zinc requirement.
• BenchTop 100bp DNA Ladder: Ready-to-load marker for agarose gel
electrophoresis. Use when gel purifying either vector or insert.
• Wizard ® SV Gel and PCR Clean-Up System: Rapid gel purification of
fragments for 100bp to 10kb. Great for removing enzymes from DNA as
well. High-capacity and low elution volume.
• PureYield Plasmid Miniprep System: Rapid 10-minute miniprep.
Prepare your vector for subcloning or use to screen for recombinants. Go
from screening to transfection thanks to the high-quality DNA.
Storage Conditions: Store the LigaFast Rapid DNA Ligation System
(M8221) and TSAP Thermosensitive Alkaline Phosphatase (M9910) at –20°C.
Store the BenchTop 100bp DNA Ladder at 22–25°C; storage at –20°C can
enhance the shelf life of this product. Store the Wizard ® SV Gel and PCR
Clean-Up System (A9281) and PureYield Plasmid Miniprep System (A1223)
at 22–25°C.
Unlike site-specific recombination vector systems, the Flexi ® Vector Systems do
not require appending multiple amino acids to the amino or carboxy termini of
the protein of interest. In addition, the systems do not require an archival entry
vector, and most applications allow direct entry into the vector suited to the
experimental design.
C-terminal Flexi ® Vectors allow expression of C-terminal-tagged proteins.
While these vectors can act as acceptors of protein-coding regions flanked by
SgfI and PmeI, they lack a PmeI site and contain a different blunt-ended site,
EcoICRI. This joined sequence cannot be removed from the C-terminal Flexi ®
Vectors and transferred to other Flexi ® Vectors.
Features:
• Versatility: You can choose between a variety of initial applications (e.g.,
bacterial protein, mammalian, or cell-free protein expression) and then
transfer to others as required.
• Time Savings: Efficient transfer allows for direct use of recombinant
clones, minimizing time wasted screening background colonies.
• Enhanced Productivity: Adaptable to high-throughput formats for large
screening projects.
• Easy Access: No licensing fees or complicated transfer restrictions.
Storage Conditions: Cat.# C8640 is comprised of Cat.# C8641 and A9280.
Store Cat.# C8641 at –20°C; store Cat.# A9280 at room temperature. Store
Cat.# C8820 and C9320 at –20°C. Store enzyme blends at –20°C.
Protocol
Flexi ® Vector Systems Technical Manual
SgfI
SgfI
protein-coding region
PCR
PmeI
DNA Purification
PmeI
Part#
TM254
5
Cloning and DNA Markers
Flexi ® Cloning System
Product Size Cat.# Price ($)
Flexi ® System, Entry/ 5 entry and 20 transfer reactions C8640 435
Transfer
Flexi ® System, Transfer 100 transfer reactions C8820 1730
Carboxy Flexi ® System,
50 transfer reactions C9320 758
Transfer
Available Separately Size Cat.# Price ($)
10X Flexi ® Enzyme Blend (SgfI & PmeI) 25 μl R1851 166
100 μl R1852 585
Carboxy Flexi ® Enzyme Blend (SgfI & EcoICRI) 50 μl R1901 165
HaloTag ® Cloning Starter System 1 each G6050 580
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The Flexi ® Vector System is a directional cloning method for
protein-coding sequences. It is based on two rare-cutting restriction enzymes,
SgfI and PmeI, and provides a rapid, efficient and high-fidelity way to transfer
protein-coding regions without the need to resequence.
All Flexi ® Vectors carry the lethal barnase gene, which is replaced by the DNA
fragment of interest and acts as a positive selection for the successful ligation
of the insert.
SgfI
Ligation
Transformation
and Selection
Digestion with Flexi ®
Enzyme Blend
DNA Purification
protein-coding region
SgfI
PmeI
lethal gene
PmeI
SgfI
lethal gene
Acceptor
Flexi ®
Vector
Digestion with Flexi ®
Enzyme Blend
PmeI
4599MA
Section
Contents
For complete and up-to-date product information visit: www.promega.com/catalog
113
Table of
Contents