Modifying Enzymes
Proteinase K
Quick Stick Ligase
T4 DNA Ligase
T4 Polynucleotide Kinase
Uracil-DNA-Glycosilase (UDGase)
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| PRODUCT |
SIZE
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CONC.
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CAT NO.
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QUANTITY
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Proteinase K |
100mg
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200-300u/mg
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BIO-37037
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1
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Proteinase K |
500mg
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200-300u/mg
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BIO-37038
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1
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Proteinase K |
1000mg
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200-300u/mg
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BIO-37039
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1
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- Features
Broad-spectrum serine
protease, useful for general
digestion of proteins.
Active in a wide pH range,
SDS, EDTA and Urea.
Stable at high
temperatures.
Molecular grade and free
of DNase and RNase activity.
- Description
Proteinase K is an enzyme
used as a protein denaturant
in many molecular-biological
techniques. The enzyme may
be used at 56ΊC for up to 4
hours, or 37ΊC for overnight
incubations.
- Applications
Inactivation of RNases and
DNases during nucleic acid
extraction.
Protein modification
- Storage Conditions
The product can be stored
for 6 months at -20ΊC in a
constant-temperature
freezer.
- Unit Definition
One unit catalyzes the
liberation in 1 minute of
Folin-positive amino acids
and peptides corresponding
to 1΅mol tyrosine.
- Specific Activity
200-300u/mg protein at 25°C
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| PRODUCT |
SIZE
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CONC.
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CAT NO.
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QUANTITY
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Quick Stick Ligase |
50 reaction
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BIO-27027
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1
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Quick Stick Ligase |
100 reactions
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BIO-27028
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1
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- Features
Rapid ligations of
cohesive and blunt-ended DNA
fragments in 5 minutes at
room temperature.
Dramatically decreases the
time required for DNA
cloning.
No loss of transformation
efficiency.
- Figure 1:
-
 Lanes
M: HyperLadder I Marker
1: Hind III-digested Lambda
DNA
2: Hind III-digested Lambda
DNA Ligated with Quick Stick
Ligase
3: EcoR V-digested Lambda
DNA
4: EcoR V-digested Lambda
DNA ligated with Quick-Stick
Kit
Samples run on 1.0% TAE
agarose gel.
Figure 1: Ligation of sticky
or blunt-ended fragments
with Bioline Quick Stick
Ligase. Lambda DNA was 10x
over-digested with EcoR V,
or Hind III, followed by
heat inactivation. DNA
fragments were ligated using
the Bioline protocol for 5
minutes at room temperature.
- Description
Quick-Stick Ligase is a T4
DNA ligase, which has been
mutated to dramatically
improve enzyme activity,
delivering fast, efficient
ligation at room
temperature. The enzyme
enables the rapid ligation
of two strands of DNA
between the 5- phosphate
and the 3- hydroxyl groups
of adjacent nucleotides in
either a blunt-ended or
cohesive-ended
configuration. Within 5
minutes, Quick Stick Ligase
will provide nearly 100%
ligation of cohesive-ended
fragments of 100ng of Hind
III- cut lambda DNA, or 80%
ligation of blunt-ended
fragments for the same
amount of EcoRV-cut lambda
DNA. 100% ligation of
blunt-ended fragments can be
achieved by increasing
ligation time to 15 minutes
at room temperature.
- Applications
Cloning of DNA from most
sources, including PCR
fragments, plasmids,
cosmids, genomic, phage and
viral DNA.
Linker ligation.
Re-ligation of linearised
plasmids.
Ligation of
double-stranded
oligonucleotides into
vectors (plasmid and phage).
- Storage Conditions
The product can be stored
for 6 months at -20ΊC in a
constant-temperature
freezer.
- Schematic
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| PRODUCT |
SIZE
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CONC.
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CAT NO.
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QUANTITY
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T4 DNA Ligase |
500u
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BIO-27026
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1
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- Features
Catalyses the joining of
two strands of DNA between
the 5- phosphate and
3-hydroxyl groups of
adjacent nucleotides.
Exhibits high activity.
- Description
High Activity.T4 DNA
Ligase catalyses the
formation of phosphodiester
bonds in the presence of ATP
between double-stranded DNAs
with 3-hydroxyl and
5-phosphate termini. The
enzyme also catalyses the
joining of RNA either to DNA
or to RNA strands in a
duplex molecule, but will
not join single-stranded
nucleic acids.
- Applications
Ligation of cohesive- and
blunt- ended DNA fragments
for cloning.
Sealing nicks in
double-stranded DNA.
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| PRODUCT |
SIZE
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CONC.
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CAT NO.
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QUANTITY
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T4 Polynucleotide Kinase |
500u
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BIO-27031
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1
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- Features
Catalyses the transfer of
the g-phosphate from ATP to
the 5-terminus of
polynucleotides or to
mononucleotides bearing a
3-phosphate group.
- Description
T4 Polynucleotide Kinase is
purified from recombinant
E.coli strain JM109, which
overproduces T4 PNK. The
enzyme is widely used to
end-label short
oligonucleotide probes and
both DNA and RNA molecules
for sequence determination.
- Applications
5 labelling of
oligonucleotides for use as
probes and sequencing
primers.
5 labelling of
single-stranded DNA and RNA
molecules for use as probes.
Sequencing of DNA protein
footprinting.
End-labelling of DNA and
RNA molecules for sequence
determination.
- Storage Conditions
The product can be stored
for 6 months at -20ΊC in a
constant-temperature
freezer.
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| PRODUCT |
SIZE
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CONC.
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CAT NO.
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QUANTITY
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Uracil-DNA-Glycosilase (UDGase) |
500u
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1u/΅l
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BIO-27044
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1
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- Features
-
- Removes carryover
contaminations.
- Recombinant source.
- Free of DNase and
RNase activities.
- Supplied with 10x
Reaction Buffer.
- Source
E.coli carrying an
overexpressed gene of
Uracil-DNA-Glycosilase from
E.coli.
- Description
Removal of Uracil from DNA.
Uracil-DNA-Glycosilase
(UDGase) catalyses the
release of Uracil from
single- or double-stranded
DNA containing Uracil, but
not from RNA. Removes
carryover contaminations
that can lead to false
positives during DNA
amplification.
- Applications
All kinds of PCR where
preventing the carryover of
contaminations is crucial,
including RT-PCR, Real Time
PCR and Site-directed
mutagenesis.
- Unit Definition
One unit is the amount of
enzyme that catalyses the
release of 60΅mol of uracil
per minute from
double-stranded DNA
containing Uracil.
- Reaction Buffers
10x Reaction Buffer:
0.25M Tris-Cl, 1mM EDTA,
10mM DTT, pH8.0. Incubate at
37°C.
- Storage Buffer
20mM Tris-Cl, pH8.0, 0.1M
NaCl, 0.1mM EDTA, 1mM DTT,
0.1% Triton X-100, 50%
glycerol.
- Storage Conditions
The product can be stored
for 6 months at -20°C in a
constant-temperature
freezer.
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+32 (0) 16
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