Download Technology Offer Use of type IIB restriction endonucleases in next

Technology Offer
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Use of type IIB restriction endonucleases in next
generation sequencing (NGS) applications
Background
Next generation sequencing (NGS) allows for a highthroughput retrieval of genomic information. The use of short
sequence reads not only enables whole genome (re)sequencing but also facilitates tasks where only short yet a
plethora of spots covering genomic information are of
importance. Multiplexing in combination with reduction of an
individual’s genomic complexity therefore allows for highthroughput genotyping, identification and comparison of
individuals or BAC-clone library mapping. In the provided
method a simple reduction of complexity is achieved by the
use of type IIB restriction endonucleases.
Max-Planck-Innovation GmbH
Amalienstr. 33
80799 Munich
Germany
Phone: +49 (89) 29 09 19 - 0
Fax:
+49 (89) 29 09 19 - 99
[email protected]
www.max-planck-innovation.de
Contact
Dr. Dieter Link
Phone: +49 (89) 29 09 19 - 28
[email protected]
Technology
Class IIB restriction endonucleases belong to a family of
enzymes that cleave DNA on both sites of their recognition
sequence leading to the release of short dsDNA (typically 3038 base pairs). Besides the recognition site of the class IIB
enzyme, the released duplex includes a unique sequence
that can be assigned to a specific region of the underlying
genome. As the read length of next generation sequencers is
still limited to a few 100s of base pairs, class IIB digested
DNA provides unique markers falling entirely within the scope
of present NGS applications. The use of these class IIB
dependent markers offers a fast and cheap method for
sequence-based physical BAC-clone library mapping, whole
genome assembly and identification of individual organisms.
We are looking for industry partners willing to enter into
licensing discussions regarding the use of type IIB restriction
endonucleases in NGS applications.
Patent Information
An international patent application has been filed that already
entered regional/national phases in Europe/US.
Figure 1. The procedure achieves the production of (barcoded) Type IIB restriction enzyme fragments ready for 2nd generation sequencing - in 3 steps, which can be performed consecutively within the same
reaction tube without intermediate cleanup. The procedure is depicted in Figure 1. (It illustrates the details
for Type IIB enzymes BcgI or AlfI, which yield 36-bp fragments with 2-bp long 3’overhangs of unknown
sequence.) The procedure will generally work for Type IIB enzymes with only minor modifications to the
adaptors, which are necessary to accommodate the different 3’ overhangs.