scholarly journals Characterization of Biomphalaria orbignyi, Biomphalaria peregrina and Biomphalaria oligoza by polymerase chain reaction and restriction enzyme digestion of the internal transcribed spacer region of the RNA ribosomal gene

2000 ◽  
Vol 95 (6) ◽  
pp. 807-814 ◽  
Author(s):  
Linus Spatz ◽  
Teofânia HDA Vidigal ◽  
Márcia CA Silva ◽  
Stella Maris Gonzalez Cappa ◽  
Omar S Carvalho
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Enzyme ◽  
Internal Transcribed Spacer ◽  
Ribosomal Gene ◽  
Enzyme Digestion ◽  
Restriction Enzyme Digestion ◽  
Chain Reaction ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain

scholarly journals Identification of four novel delta-globin gene mutations in Greek Cypriots using polymerase chain reaction and automated fluorescence- based DNA sequence analysis

Blood ◽  
1991 ◽  
Vol 78 (12) ◽  
pp. 3298-3305 ◽  
Author(s):  
P Trifillis ◽  
P Ioannou ◽  
E Schwartz ◽  
S Surrey
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Restriction Enzyme ◽  
Globin Gene ◽  
Pcr Amplification ◽  
Enzyme Digestion ◽  
Acceptor Site ◽  
Restriction Enzyme Digestion ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract The molecular basis of most beta-thalassemia syndromes has been defined, while the spectrum of mutations causing delta-thalassemia is not well characterized. In an attempt to identify such mutations, the region encompassing the delta-globin gene from three Greek Cypriot families suspected of having delta-thalassemia was amplified by polymerase chain reaction (PCR), and DNA sequence determined using an automated fluorescence-based sequencer. Four novel mutations were identified: a G----T change at codon 27 that results in an alanine to serine change; a C----T change at codon 116 converting arginine to cysteine; a T----C change at codon 141 converting leucine to proline; and an AG----GG change at the consensus 3′-acceptor site in IVS-2. While the latter is clearly a thalassemic mutation, the low hemoglobin A2 in the first three may be due to either decreased production or instability of the altered delta-globin chain. All four mutations may be detected by PCR amplification of genomic DNA followed by restriction enzyme digestion. Two mutations abolish restriction sites while two create new cleavage sites. Screening for molecular defects that cause delta-thalassemia or unstable delta-globin by PCR amplification and restriction enzyme digestion will lead to correct diagnosis of beta/delta-thalassemia compound heterozygotes and improved genetic counseling.

Diagnostic procedures and protocols.

2021 ◽  
pp. 94-142
Author(s):  
Shou-Hua Wang
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Enzyme ◽  
Diagnostic Procedures ◽  
Enzyme Digestion ◽  
Plant Diseases ◽  
Restriction Enzyme Digestion ◽  
Visual Examination ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract This chapter provides information on different diagnostic procedures and protocols for plant diseases, including visual examination, microscopy, isolation and extraction, serology-based tests, polymerase chain reaction-based detection, and restriction enzyme digestion.

scholarly journals Identification of four novel delta-globin gene mutations in Greek Cypriots using polymerase chain reaction and automated fluorescence- based DNA sequence analysis

Blood ◽  
1991 ◽  
Vol 78 (12) ◽  
pp. 3298-3305
Author(s):  
P Trifillis ◽  
P Ioannou ◽  
E Schwartz ◽  
S Surrey
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequence ◽  
Restriction Enzyme ◽  
Globin Gene ◽  
Pcr Amplification ◽  
Enzyme Digestion ◽  
Acceptor Site ◽  
Restriction Enzyme Digestion ◽  
Chain Reaction ◽  
Polymerase Chain

The molecular basis of most beta-thalassemia syndromes has been defined, while the spectrum of mutations causing delta-thalassemia is not well characterized. In an attempt to identify such mutations, the region encompassing the delta-globin gene from three Greek Cypriot families suspected of having delta-thalassemia was amplified by polymerase chain reaction (PCR), and DNA sequence determined using an automated fluorescence-based sequencer. Four novel mutations were identified: a G----T change at codon 27 that results in an alanine to serine change; a C----T change at codon 116 converting arginine to cysteine; a T----C change at codon 141 converting leucine to proline; and an AG----GG change at the consensus 3′-acceptor site in IVS-2. While the latter is clearly a thalassemic mutation, the low hemoglobin A2 in the first three may be due to either decreased production or instability of the altered delta-globin chain. All four mutations may be detected by PCR amplification of genomic DNA followed by restriction enzyme digestion. Two mutations abolish restriction sites while two create new cleavage sites. Screening for molecular defects that cause delta-thalassemia or unstable delta-globin by PCR amplification and restriction enzyme digestion will lead to correct diagnosis of beta/delta-thalassemia compound heterozygotes and improved genetic counseling.

Minimal intraspecific variation in the sequence of the transcribed spacer regions of the ribosomal DNA of lake trout (Salvelinus namaycush)

Genome ◽  
1994 ◽  
Vol 37 (4) ◽  
pp. 664-671 ◽  
Author(s):  
Lang Zhuo ◽  
S. L. Sajdak ◽  
R. B. Phillips
Keyword(s):  
Polymerase Chain Reaction ◽  
Intraspecific Variation ◽  
Ribosomal Dna ◽  
Internal Transcribed Spacer ◽  
Lake Trout ◽  
Intergenic Spacer ◽  
Coding Region ◽  
Chain Reaction ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain

Intraspecific variation in the sequence of the transcribed spacer regions of the ribosomal DNA (rDNA) in lake trout was examined by restriction mapping and sequencing of these regions amplified by the polymerase chain reaction. The length of the first internal transcribed spacer region (ITS-1) was 566 bases and the second internal transcribed spacer region (ITS-2) was 368 bases in lake trout. When the 1.4-kb region including the ITS-1, the 5.8S coding region, and the ITS-2 was amplified from 12 individuals from four populations and digested with eight different enzymes only one intraindividual polymorphism was found that occurred in each population. When the amplified ITS-1 region was sequenced from an additional 10 individuals from five populations, no interindividual variation was found in the sequence. A 6-kb portion of the rDNA repeat unit including 1.6 kb of the 18S coding region, the 5′ external spacer region (5′ ETS), and part of the adjacent intergenic spacer was cloned and a restriction map was prepared for these regions in lake trout. No intraspecific variation was found in the region adjacent to the 18S rDNA, which includes the 5′ ETS, although intraspecific and intraindividual length variation was found in the intergenic spacer region 3–6 kb from the 18S. Sequencing of a 609-b segment of the 5′ ETS adjacent to the 18S coding region revealed the presence of two 41-b repeats. The 198-b sequence between the repeats had some similarity to the 18S coding region of other fishes. Primers were designed for amplification of 559 b of the 5′ ETS using the polymerase chain reaction. No intraspecific variation in this region in lake trout was found when the DNA amplified from this region in 12 individuals from four populations was digested with eight restriction enzymes.Key words: ribosomal DNA, internal transcribed spacer regions, 5′ external spacer region, transcribed spacer, lake trout.

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