Sex determination of buffalo embryos (Bubalus bubalis) by polymerase chain reaction

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).

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Sex determination of foresic of forensic samples by polymerase chain reaction of the amelogenin gene and analysis by capillary electrophoresis with polymer matrix

Electrophoresis ◽

10.1002/elps.1150190114 ◽

1998 ◽

Vol 19 (1) ◽

pp. 76-79 ◽

Cited By ~ 8

Author(s):

Vladimir P. Pouchkarev ◽

Eugene F. Shved ◽

Peter I. Novikov

Keyword(s):

Polymerase Chain Reaction ◽

Capillary Electrophoresis ◽

Sex Determination ◽

Polymer Matrix ◽

Chain Reaction ◽

Amelogenin Gene ◽

Forensic Samples ◽

Polymerase Chain

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

Pediatrics International ◽

10.1111/j.1442-200x.1998.tb01929.x ◽

1998 ◽

Vol 40 (3) ◽

pp. 282-285

Author(s):

Yuko Jin ◽

Akihiko Endo ◽

Masami Shimada ◽

Michiyoshi Minato ◽

Masaaki Takada ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Sex Determination ◽

Chain Reaction ◽

Polymerase Chain

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Quantitative expression analysis of blastocyst-derived gene transcripts in preimplantation developmental stages of in vitro-produced bovine embryos using real-time polymerase chain reaction technology

Reproduction Fertility and Development ◽

10.1071/rd04041 ◽

2004 ◽

Vol 16 (8) ◽

pp. 753 ◽

Cited By ~ 13

Author(s):

Nermin El-Halawany ◽

Siriluck Ponsuksili ◽

Klaus Wimmers ◽

Markus Gilles ◽

Dawit Tesfaye ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Expression Pattern ◽

Embryo Development ◽

Developmental Stages ◽

Bovine Embryo ◽

Chain Reaction ◽

Quantitative Expression ◽

Polymerase Chain

The main objective of the present study was to analyse the quantitative expression pattern of genes from a subtracted blastocyst transcriptome throughout the preimplantation developmental stages of in vitro-produced bovine oocytes and embryos. For this purpose, Day 5 morula (M) cDNAs were subtracted from Day 7 blastocyst (B) cDNAs (B–M) and used to establish a B–M subtracted cDNA library, as reported previously. From the total generated clones, 19 were analysed quantitatively. The mRNA samples isolated from pools of immature oocytes (n = 150), mature oocytes (n = 150) and two-cell (n = 80), four-cell (n = 40), eight-cell (n = 20), morula (n = 6) and blastocyst (n = 3) embryos were reverse transcribed and subjected to real-time polymerase chain reaction (PCR) using sequence-specific primers and SYBR green as the DNA dye. A relative standard curve method was used to analyse the real-time data taking the morula stage as a calibrator. Applying suppression subtractive hybridisation (SSH), a total of 71 clones, which represent 33 different expressed sequence tags, were generated and available for analysis. Most transcripts were analysed for the first time in bovine embryogenesis. The real-time PCR has validated the results of SSH positively for 84% (16/19) of transcripts, whereas 16% (3/19) showed deviation in the expression pattern from the one seen during SSH. Several transcript-specific expression patterns were observed for genes that play decisive roles in bovine embryogenesis. In addition to identification, accurately quantifying the expression profiles of transcripts during development will pave the way towards understanding the molecular mechanisms of embryogenesis and their potential role in early embryo development. Most importantly, the present study has contributed to the enrichment of bovine embryo gene collection by generating new transcripts involved in bovine embryo development.

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PENENTUAN JENIS KELAMIN BURUNG KENARI (Serinus canaria) BERDASARKAN GEN Chromodomain Helicase DNA-Binding 1 (CHD1)

Jurnal Bioteknologi & Biosains Indonesia (JBBI) ◽

10.29122/jbbi.v7i1.3178 ◽

2020 ◽

Vol 7 (1) ◽

Author(s):

Afif Muhammad Akhrom ◽

Indarjulianto Soedarmanto ◽

Yanuartono Yanuartono ◽

Trini Susmiati ◽

Alfarisa Nururrozi ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Sex Determination ◽

Mature Female ◽

Mature Male ◽

Chain Reaction ◽

Blood Samples ◽

Pcr Method ◽

Polymerase Chain ◽

Female Canaries

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, sexingÂ 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.

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Frequency of Y Chromosome Microdeletions in Azoospermic and Oligospermic Iranian Infertile Men

International Journal of Medical Laboratory ◽

10.18502/ijml.v7i2.2918 ◽

2020 ◽

Author(s):

Sepideh Gholami Yarahmadi ◽

Saeid Morovvati ◽

Monireh Raam ◽

Ziba Morovvati

Keyword(s):

Polymerase Chain Reaction ◽

Y Chromosome ◽

In Vitro Fertilisation ◽

Control Group ◽

Chain Reaction ◽

Azoospermia Factor ◽

Infertile Men ◽

Y Chromosome Microdeletions ◽

Polymerase Chain

Background and Aims: Azoospermia factor (AZF) region of the Ychromosome has several genes which are responsible for normal spermatogenesis. Microdeletions of these genes are associated with azoospermia and oligospermia. These microdeletions are too small to be detected by karyotyping. They can be easily identified using polymerase chain reaction. The aim of this study is to determine the frequencies of Ychromosome microdeletions in azoospermic and oligospermic Iranian infertile men and compare them with other studies in different ethnic groups. Materials and Methods: At first, karyotype analysis was performed in 80 infertile men and 30 healthy age-matched counterparts as control group using standard cytogenetic methods. Second, genomic DNA was extracted from all cases and genetic screening was conducted for Y chromosome microdeletions by multiplex polymerase chain reaction for AZF genes on both infertile and control men using 6 STS markers on the long arm of the Y chromosome. Results: Totally, 49 infertile men were azoospermic and 31 were oligospermic. Y-chromosome microdeletions in the AZFc region were detected in 4 of azoospermic patients. Y-chromosome microdeletions was not detected in any of the oligospermic patients and the control group. Conclusions: This finding recommends that genetic counseling and screening before starting assisted reproductive techniques such as in vitro fertilisation and intracytoplasmic sperm injection can prevent unnecessary treatment and transmission of genetic defects to offspring

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