Abstract:
BACKGROUND:Splicing of DNA molecules is an important task in molecular biology that facilitates cloning, mutagenesis and creation of chimeric genes. Mutagenesis and DNA splicing techniques exist, some requiring restriction enzymes, and others utilize staggered reannealing approaches. RESULTS:A method for DNA splicing and mutagenesis without restriction enzymes is described. The method is based on mild template-dependent polymerization arrest with two molecules of cytosine arabinose (Ara-C) incorporated into PCR primers. Two rounds of PCR are employed: the first PCR produces 5' overhangs that are utilized for DNA splicing. The second PCR is based on polymerization running through the Ara-C molecules to produce the desired final product. To illustrate application of the run through stop mutagenesis and DNA splicing technique, we have carried out splicing of two segments of the human cofilin 1 gene and introduced a mutational deletion into the product. CONCLUSION:We have demonstrated the utility of a new PCR-based method for carrying out DNA splicing and mutagenesis by incorporating Ara-C into the PCR primers.
journal_name
BMC Biotechnoljournal_title
BMC biotechnologyauthors
Ailenberg M,Goldenberg NM,Silverman Mdoi
10.1186/1472-6750-5-23keywords:
subject
Has Abstractpub_date
2005-09-01 00:00:00pages
23issn
1472-6750pii
1472-6750-5-23journal_volume
5pub_type
杂志文章abstract:BACKGROUND:CRISPR/Cas9 is widely used for precise genetic editing in various organisms. CRISPR/Cas9 editing may in many plants be hampered by the presence of complex and high ploidy genomes and inefficient or poorly controlled delivery of the CRISPR/Cas9 components to gamete cells or cells with regenerative potential. ...
journal_title:BMC biotechnology
pub_type: 杂志文章
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更新日期:2019-06-17 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
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更新日期:2017-07-04 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章,收录出版
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pub_type: 杂志文章
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更新日期:2019-08-27 00:00:00
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pub_type: 杂志文章
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更新日期:2018-10-03 00:00:00
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pub_type: 杂志文章
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更新日期:2012-08-03 00:00:00
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pub_type: 杂志文章
doi:10.1186/1472-6750-7-10
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pub_type: 杂志文章
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更新日期:2018-12-14 00:00:00
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pub_type: 杂志文章
doi:10.1186/1472-6750-13-17
更新日期:2013-02-26 00:00:00
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pub_type: 杂志文章
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更新日期:2018-06-11 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:2016-04-14 00:00:00
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pub_type: 杂志文章
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更新日期:2011-01-14 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
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journal_title:BMC biotechnology
pub_type: 杂志文章
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更新日期:2006-03-06 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
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更新日期:2017-02-16 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:2011-04-07 00:00:00
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journal_title:BMC biotechnology
pub_type: 杂志文章
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更新日期:2008-09-10 00:00:00
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pub_type: 杂志文章
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更新日期:2015-05-16 00:00:00