Method for selective amplification of DNA fragments for genetic fingerprinting

The invention discloses a method for preparing a reproducible selective set of DNA fragments from the DNA or cDNA of one or more organisms as a main step in a DNA fingerprinting method. The method comprises a) cutting DNA with two or more restriction enzymes to obtain a set of double stranded DNA fragments, b) ligating to the DNA fragments double stranded adapters that match the generated fragment ends, and c) performing a polymerase chain reaction using primers which are specific for the adapters to obtain the reproducible selective set of DNA fragments. In step b) at least one modified adapter is used which is composed of two oligonucleotides that are only partly complementary, leaving at least one part of the adapter single stranded, in such a way that a complementary strand of said single stranded part of the adapter cannot be synthesized from an available 3'-end in the adapter itself, and in step c) the primer which is specific for the modified adapter, is identical or similar to the single stranded part of the adapter, such that only after complementation of the single stranded part of the adapter said primer can anneal to the complement, and in such a way that this primer can be extended in the direction of the ligation site.

Saved in:
Bibliographic Details
Main Authors: Schouten, H.J., van der Linden, C.G.
Format: Patent biblioteca
Language:English
Subjects:Life Science,
Online Access:https://research.wur.nl/en/publications/method-for-selective-amplification-of-dna-fragments-for-genetic-f
Tags: Add Tag
No Tags, Be the first to tag this record!
id dig-wur-nl-wurpubs-376535
record_format koha
spelling dig-wur-nl-wurpubs-3765352024-01-19 Schouten, H.J. van der Linden, C.G. Patent Method for selective amplification of DNA fragments for genetic fingerprinting 2004 The invention discloses a method for preparing a reproducible selective set of DNA fragments from the DNA or cDNA of one or more organisms as a main step in a DNA fingerprinting method. The method comprises a) cutting DNA with two or more restriction enzymes to obtain a set of double stranded DNA fragments, b) ligating to the DNA fragments double stranded adapters that match the generated fragment ends, and c) performing a polymerase chain reaction using primers which are specific for the adapters to obtain the reproducible selective set of DNA fragments. In step b) at least one modified adapter is used which is composed of two oligonucleotides that are only partly complementary, leaving at least one part of the adapter single stranded, in such a way that a complementary strand of said single stranded part of the adapter cannot be synthesized from an available 3'-end in the adapter itself, and in step c) the primer which is specific for the modified adapter, is identical or similar to the single stranded part of the adapter, such that only after complementation of the single stranded part of the adapter said primer can anneal to the complement, and in such a way that this primer can be extended in the direction of the ligation site. en application/pdf https://research.wur.nl/en/publications/method-for-selective-amplification-of-dna-fragments-for-genetic-f https://edepot.wur.nl/150796 Life Science Wageningen University & Research
institution WUR NL
collection DSpace
country Países bajos
countrycode NL
component Bibliográfico
access En linea
databasecode dig-wur-nl
tag biblioteca
region Europa del Oeste
libraryname WUR Library Netherlands
language English
topic Life Science
Life Science
spellingShingle Life Science
Life Science
Schouten, H.J.
van der Linden, C.G.
Method for selective amplification of DNA fragments for genetic fingerprinting
description The invention discloses a method for preparing a reproducible selective set of DNA fragments from the DNA or cDNA of one or more organisms as a main step in a DNA fingerprinting method. The method comprises a) cutting DNA with two or more restriction enzymes to obtain a set of double stranded DNA fragments, b) ligating to the DNA fragments double stranded adapters that match the generated fragment ends, and c) performing a polymerase chain reaction using primers which are specific for the adapters to obtain the reproducible selective set of DNA fragments. In step b) at least one modified adapter is used which is composed of two oligonucleotides that are only partly complementary, leaving at least one part of the adapter single stranded, in such a way that a complementary strand of said single stranded part of the adapter cannot be synthesized from an available 3'-end in the adapter itself, and in step c) the primer which is specific for the modified adapter, is identical or similar to the single stranded part of the adapter, such that only after complementation of the single stranded part of the adapter said primer can anneal to the complement, and in such a way that this primer can be extended in the direction of the ligation site.
format Patent
topic_facet Life Science
author Schouten, H.J.
van der Linden, C.G.
author_facet Schouten, H.J.
van der Linden, C.G.
author_sort Schouten, H.J.
title Method for selective amplification of DNA fragments for genetic fingerprinting
title_short Method for selective amplification of DNA fragments for genetic fingerprinting
title_full Method for selective amplification of DNA fragments for genetic fingerprinting
title_fullStr Method for selective amplification of DNA fragments for genetic fingerprinting
title_full_unstemmed Method for selective amplification of DNA fragments for genetic fingerprinting
title_sort method for selective amplification of dna fragments for genetic fingerprinting
url https://research.wur.nl/en/publications/method-for-selective-amplification-of-dna-fragments-for-genetic-f
work_keys_str_mv AT schoutenhj methodforselectiveamplificationofdnafragmentsforgeneticfingerprinting
AT vanderlindencg methodforselectiveamplificationofdnafragmentsforgeneticfingerprinting
_version_ 1813209026914156544