Chromosome Walking by Inverse PCR

2013 ◽  
pp. 299-306
Author(s):  
Michael Gotesman ◽  
Selwyn Williams ◽  
Jorge Garcés ◽  
Ray Gavin
Keyword(s):  
Inverse Pcr ◽  
Chromosome Walking

Successive Chromosome Walking by Compatible Ends Ligation Inverse PCR

2005 ◽  
Vol 30 (2) ◽  
pp. 095-102 ◽  
Author(s):  
Maozhi Ren ◽  
Quanjia Chen ◽  
Li Li ◽  
Rui Zhang ◽  
Sandui Guo
Keyword(s):  
Inverse Pcr ◽  
Chromosome Walking

Isolation of the Promoter Region of a Pollen-specific Gene PSG076 by Inverse-PCR

2012 ◽  
Vol 30 (3) ◽  
pp. 309
Author(s):  
Ling CHEN ◽  
Pei-Pei SU ◽  
Han-Wen TONG ◽  
Yi-Ke LIU ◽  
Zhan-Wang ZHU ◽  
...  
Keyword(s):  
Promoter Region ◽  
Inverse Pcr ◽  
Specific Gene

AFLP-derived, Codominant Markers for Locus-specific Applications

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 514e-514
Author(s):  
James M. Bradeen ◽  
Philipp W. Simon
Keyword(s):  
Linkage Mapping ◽  
Large Scale ◽  
Pcr Primers ◽  
Inverse Pcr ◽  
Sequence Information ◽  
Pcr Assay ◽  
Specific Primers ◽  
Simultaneous Evaluation ◽  
Feral Populations ◽  
Diversity Assessment

The amplified fragment length polymorphism (AFLP) is a powerful marker, allowing rapid and simultaneous evaluation of multiple potentially polymorphic sites. Although well-adapted to linkage mapping and diversity assessment, AFLPs are primarily dominant in nature. Dominance, relatively high cost, and technological difficulty limit use of AFLPs for marker-aided selection and other locus-specific applications. In carrot the Y2 locus conditions carotene accumulation in the root xylem. We identified AFLP fragments linked to the dominant Y2 allele and pursued conversion of those fragments to codominant, PCR-based forms useful for locus-specific applications. The short length of AFLPs (≈60 to 500 bp) precludes development of longer, more specific primers as in SCAR development. Instead, using sequence information from cloned AFLP fragments for primer design, regions outside of the original fragment were amplified by inverse PCR or ligation-mediated PCR, cloned, and sequenced. Differences in sequences associated with Y2 vs. y2 allowed development of simple PCR assays differentiating those alleles. PCR primers flanking an insertion associated with the recessive allele amplified differently sized products for the two Y2 alleles in one assay. This assay is rapid, technologically simple (requiring no radioactivity and little advanced training or equipment), reliable, inexpensive, and codominant. Our PCR assay has a variety of large scale, locus-specific applications including genotyping diverse carrot cultivars and wild and feral populations. Efforts are underway to improve upon conversion technology and to more extensively test the techniques we have developed.

scholarly journals Identification of the insertion site of transgenic DNA based on cyclization of the target gene with the flanking sequence and nested inverse PCR

Talanta Open ◽  
2021 ◽  
Vol 3 ◽  
pp. 100033
Author(s):  
Jiayu Wang ◽  
Xuetong Bi ◽  
Wei Chen ◽  
Qinyue Zhao ◽  
Jinqi Yang ◽  
...  
Keyword(s):  
Target Gene ◽  
Insertion Site ◽  
Inverse Pcr ◽  
Flanking Sequence

Identification of Chromosome Inheritance Modifiers in Drosophila melanogaster

Genetics ◽  
2001 ◽  
Vol 157 (4) ◽  
pp. 1623-1637 ◽  
Author(s):  
Kenneth W Dobie ◽  
Cameron D Kennedy ◽  
Vivienne M Velasco ◽  
Tory L McGrath ◽  
Juliani Weko ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Biological Activity ◽  
Cancer Progression ◽  
Birth Defects ◽  
Mitotic Chromosome ◽  
P Element ◽  
Inverse Pcr ◽  
Gtpase Activating Protein ◽  
New Genes ◽  
Genomic Analyses

Abstract Faithful chromosome inheritance is a fundamental biological activity and errors contribute to birth defects and cancer progression. We have performed a P-element screen in Drosophila melanogaster with the aim of identifying novel candidate genes involved in inheritance. We used a “sensitized” minichromosome substrate (J21A) to screen ∼3,000 new P-element lines for dominant effects on chromosome inheritance and recovered 78 Sensitized chromosome inheritance modifiers (Scim). Of these, 69 decreased minichromosome inheritance while 9 increased minichromosome inheritance. Fourteen mutations are lethal or semilethal when homozygous and all exhibit dramatic mitotic defects. Inverse PCR combined with genomic analyses identified P insertions within or close to genes with previously described inheritance functions, including wings apart-like (wapl), centrosomin (cnn), and pavarotti (pav). Further, lethal insertions in replication factor complex 4 (rfc4) and GTPase-activating protein 1 (Gap1) exhibit specific mitotic chromosome defects, discovering previously unknown roles for these proteins in chromosome inheritance. The majority of the lines represent mutations in previously uncharacterized loci, many of which have human homologs, and we anticipate that this collection will provide a rich source of mutations in new genes required for chromosome inheritance in metazoans.

scholarly journals Chromosome Walking by Cloning of Distinct PCR Fragments

BioTechniques ◽  
2001 ◽  
Vol 30 (2) ◽  
pp. 248-249 ◽  
Author(s):  
Bernd Kneidinger ◽  
Michael Graninger ◽  
Paul Messner
Keyword(s):  
Chromosome Walking
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