Intensive Linkage Mapping in a Wasp (Bracon hebetor) and a Mosquito (Aedes aegypti) With Single-Strand Conformation Polymorphism Analysis of Random Amplified Polymorphic DNA Markers

Genetics ◽  
1996 ◽  
Vol 143 (4) ◽  
pp. 1727-1738
Author(s):  
Michael F Antolin ◽  
Christopher F Bosio ◽  
Julie Cotton ◽  
William Sweeney ◽  
Michael R Strand ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Random Amplified Polymorphic Dna ◽  
Single Strand Conformation Polymorphism ◽  
Sscp Analysis ◽  
Agarose Gel ◽  
Linkage Maps ◽  
Agarose Gel Electrophoresis ◽  
Pcr Markers ◽  
Rapd Pcr ◽  
Conformation Polymorphism

Abstract The use of random amplified polymorphic DNA from the polymerase chain reaction (RAPD-PCR) allows efficient construction of saturated linkage maps. However, when analyzed by agarose gel electrophoresis, most RAPD-PCR markers segregate as dominant alleles, reducing the amount of linkage information obtained. We describe the use of single strand conformation polymorphism (SSCP) analysis of RAPD markers to generate linkage maps in a haplodiploid parasitic wasp Bracon (Habrobracon) hebetor and a diploid mosquito, Aedes aegypti. RAPD-SSCP analysis revealed segregation of codominant alleles at markers that appeared to segregate as dominant (band presence/band absence) markers or appeared invariant on agarose gels. Our SSCP protocol uses silver staining to detect DNA fractionated on large thin polyacrylamide gels and reveals more polymorphic markers than agarose gel electrophoresis. In B. hebetor, 79 markers were mapped with 12 RAPD primers in six weeks; in A. aegypti, 94 markers were mapped with 10 RAPD primers in five weeks. Forty-five percent of markers segregated as codominant loci in B. hebetor, while 11% segregated as codominant loci in A. aegypti. SSCP analysis of RAPD-PCR markers offers a rapid and inexpensive means of constructing intensive linkage maps of many species.

scholarly journals Detecting familial defective apolipoprotein B-100: three molecular scanning methods compared

1997 ◽  
Vol 43 (9) ◽  
pp. 1630-1634 ◽  
Author(s):  
Bryony G Henderson ◽  
Philip R Wenham ◽  
J Peter Ashby ◽  
Gillian Blundell
Keyword(s):  
Apolipoprotein B ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Single Strand Conformation Polymorphism ◽  
Sscp Analysis ◽  
Heteroduplex Analysis ◽  
Apo B ◽  
Large Numbers ◽  
Gradient Gel Electrophoresis ◽  
Conformation Polymorphism

Abstract Familial defective apolipoprotein (apo) B-100 (FDB), a condition that may give rise to hypercholesterolemia, is caused by mutations around codon 3500 of the apo B gene. We have compared the ability of three molecular-scanning techniques, heteroduplex analysis, single-strand conformation polymorphism (SSCP) analysis, and denaturing gradient gel electrophoresis (DGGE), to detect these mutations in a cohort of 432 hypercholesterolemic individuals. Heteroduplex analysis and DGGE detected 11 individuals with apo B mutations, 9 of whom were heterozygous for apo B R3500Q and 2 who were heterozygous for apo B R3531C. Whereas DGGE was able to distinguish between these two mutations, heteroduplex analysis was technically simpler and gave a higher sample throughput. In contrast, SSCP analysis detected only 7 of the R3500Q and none of the R3531C heterozygotes and was the most complex of the three techniques. We believe heteroduplex analysis to be the method of choice for screening large numbers of samples for FDB.

Luminography – an Alternative Assay for Detection of von Willebrand Factor Multimers

1988 ◽  
Vol 60 (02) ◽  
pp. 133-136 ◽  
Author(s):  
R Schneppenheim ◽  
H Plendl ◽  
U Budde
Keyword(s):  
Von Willebrand Factor ◽  
Gel Electrophoresis ◽  
Detection Methods ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Luminescence Assay ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryA luminescence assay was adapted for detection of von Willebrand factor multimers subsequent to SDS-agarose gel electrophoresis and electroblotting onto nitrocellulose. The method is as fast as chromogenic detection methods and appears to be as sensitive as autoradiography without the disadvantages of the latter.

scholarly journals Sequence Variations in the Bovine Growth Hormone Gene Characterized by Single-Strand Conformation Polymorphism (SSCP) Analysis and Their Association with Milk Production Traits in Holsteins

Genetics ◽  
1996 ◽  
Vol 144 (4) ◽  
pp. 1809-1816 ◽  
Author(s):  
Jianbo Yao ◽  
Samuel E Aggrey ◽  
David Zadworny ◽  
J Flan Hayes ◽  
Urs Kühnlein
Keyword(s):  
Growth Hormone ◽  
Milk Production ◽  
Milk Yield ◽  
Single Strand Conformation Polymorphism ◽  
Sscp Analysis ◽  
Bovine Growth Hormone ◽  
Production Traits ◽  
Milk Production Traits ◽  
Sequence Variations ◽  
Conformation Polymorphism

Sequence variations in the bovine growth hormone (GH) gene were investigated by single strand conformation polymorphism (SSCP) analysis of seven amplified fragments covering almost the entire gene (2.7 kb). SSCPs were detected in four of these fragments and a total of six polymorphisms were found in a sample of 128 Holstein bulls. Two polymorphisms, a T→C transition in the third intron (designated GH4.1) and an A→C transversion in the fifth exon (designated GH6.2), were shown to be associated with milk production traits. GH4.1c/GH4.1c bulls had higher milk yield than GH4.1c/GH4.1t (P ≤ 0.005) and GH4.1t/GH4.1t (P ≤ 0.0022) bulls. GH4.1c/GH4.1c bulls had higher kg fat (P ≤ 0.0076) and protein (P ≤ 0.0018) than GH4.1c/GH4.1t bulls. Similar effects on milk production traits with the GH6.2 polymorphism were observed with the GH6.2a allele being the favorable allele. The average effects of the gene substitution for GH4.1 and GH6.2 are similar, with ±300 kg for milk yield, ±8 kg for fat content and ±7 kg for protein content per lactation. The positive association of GH4.1c and GH6.2a with milk production traits may be useful for improving milk performance in dairy cattle.

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