Hayvan Sağlığı Araştırmaları 2009 Yılı Program - Tagem

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2. Saha ç�� çin çiftliklerden de örnekler �� ümü�� 8.3.2. Metot: Projenin laboratuar ça�� TSA ve BHIA besiyerlerine ekilerek 25-�� Lactococcus garvieae�� yürütülecek ve elde edilen bantlar jel görüntüleme sisteminde görüntülenerek uygun software ��Lactococcus garvieae�� Triptik soy agar besiyerinde saf kültür �� L. garvieae’nin tek kolon�� 600 de 0.8 -�� ve 50ºC’ye su banyosu�� Bakteriler 4°C’de, 10,000 x g de 5 dakika santrifüj edilecek ve 50ºC’lik su banyosunda pelet hücre süspansiyon tamponu (10 mM Tris, pH 7.2, 20 mM NaCl, 50 mM EDTA) ile her bir mililitrede 5 x 10 8 � �� . %2`lik agaroz ile hücre �� tüplerine 5 ml liziz tamponu (10 mM Tris, pH 7.2, 50 mM NaCl, %0.2 sodium deoxycholate, %0.5 sodium lauryl sarcosine, 1 mg/ml lysozyme) eklenecektir. Plaklar 1 saat 37ºC de inkübe �� deoxycholate, %1 sodium lauryl sarcosine, 1 mg/ml Proteinase K) eklenecek ve plaklar 50ºC’de bir gece inkübe edilecektir. Plaklar 4 defa �� lacak, tüpe 0.3 ml 1X enzim tamponu ve her bir 100 ��çin 50 U enzim eklenerek ��Lactococcus�� NotI, ApaI, XbaI, XhoI and Spe�� ndan (Fredriksson-Ahomaa ve ark., 1999) Lactococcus garvieae �� ktir. Biometra Rotaphor ile ��–��–90 V’a ayarlanarak DNA �� 186
�� (Pfaller ve ark., 1992; Hollis ve ark., 1999�� L. garvieae’nin �� Lactococcus garvieae’nin genotiplerinin belirlenmesinde kulla�� edilen L. garvieae�� 9. Ö�� mevcuttur. �� Veteriner hekim, SUMAE de 2004 �� 10.1. Birim�� bilgi birikimine sahiptir. 10.2. �� : Pulsed-Field Jel Elektroforez (PFGE) �� 10.3. �� Etkinlikleri: Enstitümüzde Su ��–�� 187
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Hayvan Sağlığı Araştırmaları 2009 Yılı Program - Tagem

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