Tn5 Transposase

$490.00$1,990.00

MCLAB’s Tn5 Transposase represents a highly active variant of the Tn5 transposase enzyme. This enzyme facilitates the random insertion of a Tn5 Transposon into target DNA in vitro.

SKU: N/A Category:
SKU Sizes PriceQuantity
TN5-100 100µL (10 pmol/μl) $490.00
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TN5-200 500µL (10 pmol/μl) $1,990.00
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Description

Description:

MCLAB’s Tn5 Transposase represents a highly active variant of the Tn5 transposase enzyme. This enzyme facilitates the random insertion of a Tn5 Transposon into target DNA in vitro. Efficient transposition requires the presence of a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each end of the Tn5 Transposon.

The transposition process catalyzed by the Tn5 Transposase involves a multi-step “cut and paste” mechanism. Initially, the enzyme binds to the 19-bp ME of the transposon, forming a complex known as the Transposome. Subsequently, the Transposome initiates random attacks on the phosphodiester backbone of the target DNA, leading to cleavage. Finally, the Tn5 Transposase catalyzes the covalent linkage of the 3′-OH ends of the transposon to the exposed 5′-phosphorylated ends of the target DNA. This transposition event results in the creation of a 9-bp sequence duplication immediately flanking the site of transposon insertion.

Applications:

  • Perform an in vitro transgenic experiment
  • Construct a random library for second-generation sequencing
  • Incorporate a marker, such as a T7 promoter or R6Kγ origin of replication, into any desired target DNA.
  • Integrate any custom DNA sequence, bordered by one or both ends of the 19-bp MEs of an Tn5 Transposon, into a chosen target DNA.
  • Prepare Transposomes without Mg2+ for electroporation into live bacteria, enabling the subsequent random insertion of the transposon into the bacterial chromosome

Source:

Purified from an E. coli strain carrying the Tn5 gene.

MCLAB # Component Name Concentration Storage Conditions Amount
TN5-100
Tn5 Transposase 1 U/µL -20°C 100 µL 
10X Tn5 Reaction Buffer 10X -20°C 1×500µl
10X Tn5 Stop Solution 10X -20°C 1×500µl
TN5-500
Tn5 Transposase 1 U/µL -20°C 500 µL
10X Tn5 Reaction Buffer 10X -20°C 1x1ml
10X Tn5 Stop Solution 10X -20°C 1x1ml

Unit Definition:

One unit of Tn5 Transposase catalyzes the release of the donor backbone fragment from 1 μg of transposed DNA in 1 hour at 37°C, as determined by agarose gel electrophoresis.

Supplied with:

10X Tn5 Transposase Reaction Buffer
10XTn5 Stop Solution

10X Tn5 Transposase Reaction Buffer
1 M Tris-acetate (pH 7.5)
3 M potassium acetate
200 mM magnesium acetate

10X Tn5 Stop Solution
1% SDS

Supplied in:

Storage Buffer:
50% glycerol
50 mM Tris-HCl (pH 7.5)
100 mM NaCl
0.1 mM EDTA
0.1% Triton® X-100 (Rohm & Haas)
1 mM dithiothreitol

Recommended Storage Condition: -20°C, avoid repeated free-thaw

Additional information

Sizes

100µL (10 pmol/μl), 500µL (10 pmol/μl)

Manual & Protocols

The Q&A for this product will be available soon.