Selected research plan

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IVEP in Cattle

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Selected research plan:

M5430154 MS.Thabtim Samdangchai D5430024 MS.Tayita Suttirojpattana D5430031 Mr.Theesit Juanpanich D5430246 Mr.Ashit Kumar Paul 1 Selected research plan

Supervisor:

Supervisor 2 Dr. Rungsun Parnpai Associate Professor School of biotechnology Institute of Agricultural Technology Suranaree University of Technology

Research plan:

Research plan 3 Oocyte collection IVM Oocyte fixation and staining IVF IVC Vitrification Vitrification

Oocyte collection:

Oocyte collection 4 Slaughter house 35 o C, 0.9% NaCl solution Aspirated follicle from bovine ovary with 18 –gauge needle attached to 10 ml syringe

In vitro maturation (IVM):

In vitro maturation (IVM) 5 Cultured in TCM 199 medium Incubate for 24 hours in 5%CO 2 in humidified air at 39 o C

In vitro fertilization (IVF):

In vitro fertilization (IVF) 6 Sperm preparation two straw thawed at 37 o C for 30 seconds. the thawed semen will be layered on Percoll medium centrifuged at 900 rpm for 15 minutes In vitro insemination 44 u l drop of IVF-TL incubate for 2 hours. 10 COCs will be transfer to IVF-TL and co-incubated for 24 hours in 5%CO 2 in humidified air at 39 o C.

In vitro culture (IVC) :

In vitro culture (IVC) 7 After co-incubation, vortexed for 1-2 minute and culture in IVC medium for next 135 hours 5%CO 2 in humidified air at 39 o C for blastocyst.

Vitrification:

Vitrification 8 Vitrification means cooling without formation of ice. It is the process to preserve the oocyte or embryo by liquid nitrogen for long time. Storage in LN 2

Objectives of this plan:

Objectives of this plan To investigate the In vitro production of embryo in cattle To develop the high productive cattle breed To bypass the dependency of whole follicular development in cattle To increase the livestock population

PowerPoint Presentation:

Work Plan 1. Oocytes manipulation (1st and 2nd week) (9th Jan to 22 Jan) 2. In vitro maturation (3rd and 4th week) (23 Jan-5th Feb.) 3. In vitro fertilization and embryo culture (5th and 6th week) (6 feb-19 Feb) 4. Vitrification of oocytes (9th and 10th week) (5 march- 18march) 5. Vitrification of embryos (11st and 12nd week) (19Mrch-1april) 6. Embryo Transfer (11th and 12th week) (2april-15april)

PowerPoint Presentation:

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