scholarly journals Identification of the GDF9 mutation in two sheep breeds by using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) technique

2010 ◽  
Vol 9 (47) ◽  
pp. 8020-8022 ◽  
Author(s):  
Ghaderi A ◽  
T Beigi Nasiri M ◽  
H Mirzadeh K ◽  
Fayazi J ◽  
S Sadr A
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

scholarly journals Análise de polimorfismo do gene da somatotropina em vacas Nelore e seu efeito sobre o peso à desmama de suas progênies

1999 ◽  
Vol 51 (6) ◽  
pp. 565-570
Author(s):  
F.J.C. Faria ◽  
S.E.F. Guimarães ◽  
R.M.G. Lima ◽  
G.B. Mourão ◽  
L.E.L. Pinheiro
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Informações sobre peso à desmama de um rebanho Nelore foram utilizadas após ajuste para idade padrão de 205 dias, sexo da cria, idade da mãe, touro e mês de desmama, para separar as reprodutrizes em dois grupos, cujos filhos diferiam nesse peso. As médias ajustadas pelo método dos quadrados mínimos foram para os grupos pesado (P) e leve (L) de 163,21± 2,18kg e 134,44± 2,18kg, respectivamente, com 41 animais em cada grupo. Essas reprodutrizes foram submetidas à coleta de sangue para estudo de polimorfismos do gene da somatotropina bovina, pela técnica de PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism). A amplificação de uma região entre o éxon III e V do gene da somatotropina permitiu analisar dois sítios de restrição. Para o sítio do éxon V, todos os animais foram identificados como monomórficos (Leu-Leu). Quanto ao sítio do íntron 3, foi possível identificar os seguintes genótipos 21 (+/-) e 60 (-/-), com as freqüências de 0,13 e 0,87 para os alelos (+) e (-), respectivamente. O peso dos filhos dos animais com o genótipo +/- foi de 152,42± 4,41kg e os -/- 147,60± 2,61kg. Os grupos P e L não diferiram entre si quanto às freqüências alélicas apresentadas. O genótipo das reprodutrizes não afetou o peso à desmama de suas crias, existindo portanto outros efeitos genéticos e não genéticos de maior magnitude.

scholarly journals Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Authentication of Raw Meats from Game Birds

2008 ◽  
Vol 91 (6) ◽  
pp. 1416-1422 ◽  
Author(s):  
María Rojas ◽  
Isabel González ◽  
Violeta Fajardo ◽  
Irene Martín ◽  
Pablo E Hernández ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Abstract Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis has been applied to the identification of meats from quail (Coturnix coturnix), pheasant (Phasianus colchicus), red-legged partridge (Alectoris rufa), guinea fowl (Numida meleagris), capercaillie (Tetrao urogallus), Eurasian woodcock (Scolopax rusticola), woodpigeon (Columba palumbus), and song thrush (Turdus philomelos). PCR amplification was performed using a set of primers flanking a conserved region of approximately 720 base pairs (bp) from the mitochondrial 12S rRNA gene. Restriction site analysis based on sequence data from this DNA fragment permitted the selection of Alul and BfaI endonucleases for species identification. The restriction profiles obtained when amplicons were digested with the chosen enzymes allowed the unequivocal identification of all game bird species analyzed. However, the use of the PCR-RFLP technique described is limited to raw meat authentication. It is not suitable for cooked products because thermal treatment strongly accelerates DNA degradation leading to difficulties in amplifying the 720 bp fragment.

scholarly journals Προσδιορισμός της συμβολής υπόπτων γονιδίων υπευθύνων για την ηλικιακή εκφύλιση της ωχράς κηλίδας στην Ελλάδα

2015 ◽  
Author(s):  
Ιωσήφ Σκαλιδάκης
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

ΣΚΟΠΟΣ : Αξιολόγηση της συμβολής των πολυμορφισμών στα γονίδια CFH ,ARMS2, APOE ,FCGR2A στην προδιάθεση για εκδήλωση ΗΕΩ (ηλικιακής εκφύλισης ωχράς κηλίδας) στον Ελληνικό πληθυσμό.ΜΕΘΟΔΟΛΟΓΙΑ : Η απομόνωση του DNA πραγματοποιήθηκε με στήλες φυγοκέντρησης QIAamp,από αίμα ασθενών με ΗΕΩ και υγιών (κατόπιν οφθαλμολογικού ελέγχου). Για τη γονοτύπωση των πολυμορφισμών στα γονίδια ARMS2 και APOE χρησιμοποιήθηκε η τεχνική PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism) και επιπλέον αναπτύχθηκε νέα μεθοδολογία με PCR πραγματικού χρόνου και ανάλυση καμπύλων τήξης για τον πολυμορφισμό Y402H του γονιδίου CFH και H131R του FCGR2A. Ακολούθησε στατιστική ανάλυση με προγράμματα SNPStats και SPSS .ΑΠΟΤΕΛΕΣΜΑΤΑ : Πραγματοποιήθηκε αρχικά έλεγχος για συσχετισμό των 4 γονιδίων αυτών με την ΗΕΩ σε δείγματα αίματος 27 πασχόντων και 34 υγιών εθελοντών αναδεικνύοντας μια ισχυρή συσχέτηση της νόσου με τους Υ402Η στο CFH (OR=2,68) και H131R στο FCGR2A (OR=3.17) ενώ δεν υπήρξε στατιστικά σημαντική συσχέτηση της νόσου με τους πολυμορφισμούς σε ARMS2 και ApoE . Σε δεύτερο χρόνο μελετήθηκε σε δείγματα 120 ασθενών και 103 υγιών η συμβολή των πολυμορφισμών στα CFH και FCGR2A για εκδήλωση ΗΕΩ όπου φάνηκε να υπάρχει στατιστικά σημαντική συσχέτηση με OR=1,77 και 1,74 αντίστοιχα. ΣΥΜΠΕΡΑΣΜΑ : Η μελέτη για πρώτη φορά της συσχέτισης της ΗΕΩ με τον πολυμορφισμό Η131R απέδωσε σημαντικά αποτελέσματα, προτείνοντας ισχυρή συσχέτιση και μεγάλο σχετικό κίνδυνο για τους φορείς της μετάλλαξης.

Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) for the identification of Trichinella isolates

Parasitology ◽  
1999 ◽  
Vol 118 (2) ◽  
pp. 211-218 ◽  
Author(s):  
Z. WU ◽  
I. NAGANO ◽  
E. POZIO ◽  
Y. TAKAHASHI
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

In the present study, polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis was developed to identify 5 species (Trichinella spiralis, Trichinella britovi, Trichinella nativa, Trichinella nelsoni and Trichinella pseudospiralis) and 3 phenotypes of uncertain taxonomic status (Trichinella T5, T6, and T8). Eleven restriction endonucleases were used to restrict 3 DNA fragments (1) a 2800 bp fragment of the 43 kDa excretory–secretory (E–S) protein gene, (2) a 1250 bp fragment amplified with the primer pair SB147A and (3) a 372 bp fragment amplified with the primer pair SB372A. This RFLP method allows the identification of the 8 Trichinella phenotypes as follows: T. spiralis by the HinfI or DdeI endonuclease restriction of the 2800 bp fragment; T. nativa by the RsaI restriction of the 2800 bp fragment, or by the AluI restriction of the 1250 bp fragment; T. britovi and Trichinella T8 by the AluI restriction of the 1250 bp fragments, and can be discriminated between them by the SspI restriction of the 2800 bp fragment; T. pseudospiralis by the MspI restriction of the 372 bp fragment; T. nelsoni by the HhaI or AluI restriction of the 2800 bp fragment; Trichinella T5 by the HhaI restriction of the 2800 bp fragment; Trichinella T6 by the AluI restriction of the 1250 bp fragment; and Trichinella T8 by the SspI or RsaI restriction of the 2800 bp fragment. This study reveals also an intraspecifies polymorphism in the 2800 bp and 1250 bp fragments for T. britovi, Trichinella T5 and T6.

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