EZ-Tn5™ <oriV/KAN-2> Insertion Kit

Catalogue number

Pack size

EZI02VK

10 reactions

EZ-Tn5™ Transposase

Catalogue number

Pack size

TNP92110

10U

TransforMax™ EPI300™-T1^R Electrocompetent E. coli

Catalogue number

Pack size

EC02T110

10 x 100µl

EC02T15

5 x 100µl

TransforMax™ EPI300v Electrocompetent E. coli

Catalogue number

Pack size

EC300105

5 x 100µl

EC300110

10 x 100µl

EC300150

50 x 100µl

Insert Priming Sites and Convert Single-Copy Clones to Multi-Copy-Inducible

The EZ-Tn5™ <oriV/KAN-2> Insertion Kit is designed to simplify and speed up both DNA purification and sequencing of existing single-copy BAC and fosmid clones by integrating CopyControl™ capability. Use a short, one-step in vitro reaction to randomly insert a single EZ-Tn5™ <oriV/KAN-2> Transposon containing an inducible, high-copy origin of replication (oriV) into the target DNA. Then, transform TransforMax™ EPI300™ or EPI300v-T1^R Electrocompetent E. coli (sold separately) with an aliquot of the reaction and select for the kanamycin marker encoded by the EZ-Tn5™ Transposon.

One reaction will generate thousands of EZ-Tn5 <oriV/KAN-2> Transposon insertion clones. Each clone not only contains oriV and a kanamycin marker, but primer binding sites at the ends of the inserted transposon for sequencing. Amplify selected clones by adding the CopyControl™ Induction Solution and purify microgram quantities of template DNA from as little as 1ml of culture. Use the primers provided in the kit to sequence bidirectionally from the ends of the transposon and save the time and money usually spent subcloning or making custom primers.

Figure 1. The process for generating EZ-Tn5™ <oriV/KAN-2> Transposon insertion clones for high yields of DNA and bidirectional sequencing.

Last modified: 2008-05-14 16:46:46

Benefits

• Make single-copy BAC and fosmid clones inducible to high-copy number for higher yields of higher purity DNA
• A single reaction generates thousands of independent insertion clones, each with a single EZ-Tn5™ Transposon at a different site
• Completely sequence BAC and fosmid clones with only two sequencing primers and avoid shotgun subcloning or primer walking

Last modified: 2008-05-14 16:46:46

Protocols for: EZ-Tn5™ <oriV/KAN-2> Kits

EZ-Tn5™ <oriV/KAN-2> Kit Protocol

(catalogue number EZI02VK / URP023 / UFP024)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46