Using polymerase chain reaction for rapid sex-determination of a case of sirenomielia

1998 ◽  
Vol 40 (3) ◽  
pp. 282-285
Author(s):  
Yuko Jin ◽  
Akihiko Endo ◽  
Masami Shimada ◽  
Michiyoshi Minato ◽  
Masaaki Takada ◽  
...  
Author(s):  
Afif Muhammad Akhrom ◽  
Indarjulianto Soedarmanto ◽  
Yanuartono Yanuartono ◽  
Trini Susmiati ◽  
Alfarisa Nururrozi ◽  
...  

Phenotype determination of sex in young canaries is very low in accuracy. This study aimed to develop a genotypic sexing method in canaries. This study used 12 canaries consisting of 3 mature males, 3 mature females and 6 one-month-old canaries. Phenotypic sexing by cloacal observation was done on all birds, continued by genotypic sexing to identification CHD1 gene using polymerase chain reaction (PCR). The PCR used blood samples for mature canaries, and feather for mature and one-month-old canaries. The results of phenotypic observations showed that all mature male canaries had prominent and pointed cloaca forms, all mature females had flat and wide, whereas all one-month-old birds had a flat cloaca. The result of PCR showed a single band (500 bp) for mature male and double bands (500 bp and 300 bp) for mature female canaries. The PCR results of one-month-old canaries showed that there were one male and five females. Based on this study, it was concluded that genotypic sexing using the PCR method is effective in the sex determination of canaries.Keywords: canary, CHD1, genotype, PCR, sexing ABSTRAKPenentuan jenis kelamin burung kenari muda secara fenotip akurasinya sangat rendah. Penelitian ini bertujuan untuk menentukan jenis kelamin burung kenari secara genotip. Penelitian ini menggunakan 12 ekor burung kenari, terdiri dari 6 ekor dewasa (3 jantan, 3 betina) serta 6 ekor umur 1 bulan. Semua burung ditentukan jenis kelaminnya dengan mengamati kloaka dan identifikasi gen CHD1 menggunakan teknik polymerase chain reaction (PCR). Sampel DNA berasal dari darah dan bulu untuk burung dewasa serta bulu untuk burung umur 1 bulan. Pengamatan fenotip menunjukkan bahwa burung kenari dewasa jantan mempunyai bentuk kloaka menonjol dan runcing, dewasa betina berbentuk datar dan lebar, sedangkan semua burung umur 1 bulan mempunya bentuk kloaka datar. Hasil identifikasi gen CHD1 diperoleh adanya 1 pita gen sekitar 500 bp dari sampel darah dan bulu semua burung kenari dewasa jantan, dan 2 pita gen sekitar 500 bp dan 300 bp dari sampel semua burung kenari betina dewasa. Hasil PCR pada sampel burung umur 1 bulan menunjukkan bahwa 1 ekor jantan dan 5 ekor betina. Berdasarkan penelitian ini dapat disimpulkan bahwa penentuan jenis kelamin secara genotip menggunakan gen CHD1 dapat dilakukan pada burung kenari.


Zygote ◽  
2003 ◽  
Vol 11 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Laura Manna ◽  
Gianluca Neglia ◽  
Marcella Marino ◽  
Bianca Gasparrini ◽  
Rossella Di Palo ◽  
...  

The aim of this study was to identify a simple, rapid method for sex determination of in vitro produced buffalo embryos, amplifying Y-chromosome-specific repeat sequences by polymerase chain reaction (PCR). Buffalo oocytes collected from slaughtered animals were matured, fertilised and cultured in vitro for 7 days. On day 7 embryos were evaluated and divided in to six groups according to developmental stage (2, 4, 8, 16 cells, morulae and blastocyst). Each embryo was stored singly in phosphate-buffered saline at −20 °C until PCR. Two different methods of extraction of DNA were compared: a standard procedure (ST), using a normal extraction by phenol-chloroform, isoamyl alcohol and final precipitation in absolute ethanol and a direct procedure (DT), using a commercial kit (Qiaquik-Qiagen mini blood). A pair of bovine satellite primers and two pairs of different bovine Y-chromosome-specific primers (BRY4.a and BRY.1) were used in the PCR assay on embryos and on whole blood samples collected from male and female adult buffaloes, used as control. The trial was carried out on 359 embryos (193 for ST and 166 for DT). When DNA samples from blood were amplified, the sex determined by PCR always corresponded to the anatomical sex. Embryo sexing was not possible in two embryos in ST and one embryo in DT. Both extraction protocols recovered sufficient quantities of target DNA at all developmental stages, but the time required for the ST (24 h) limits its use in embryo sexing and supports the use of commercial extraction kits (5 h).


2013 ◽  
Vol 48 (6) ◽  
pp. 1001-1005 ◽  
Author(s):  
K Kaneko ◽  
E Uematsu ◽  
Y Takahashi ◽  
B Tong ◽  
S Takino ◽  
...  

1990 ◽  
Vol 1 (2) ◽  
pp. 121-133 ◽  
Author(s):  
A. Schröder ◽  
J. R. Miller ◽  
P. D. Thomsen ◽  
K. Roschlau ◽  
B. Avery ◽  
...  

1995 ◽  
Vol 1 (1) ◽  
pp. 45-50
Author(s):  
J.C.F.M. Dreesen ◽  
J.C.M. Dumoulin ◽  
J.L.H. Evers ◽  
J.P.M. Geraedts ◽  
M.H.E.C. Pieters

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