Tn5 Transposase

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.

Enzyme is supplied at a concentration of 1 U/µL.

Application:

• 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.

• Storage:

-20°C, avoid repeated free-thaw

• Supplied with:

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)

• Size:

100 µL (cat.n. TN5-100)

500 µL (cat.n. TN5-200)