NDRI Quinquennium Book COVER FINAL CURVE.cdr

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Cloned calf (Garima-II) using embryonic stem cell as donorwas born on August 22, 2010.A cloned calf was produced from a frozen-thawed clonedembryo. This male calf christened ‘SHRESTH’ was born onAugust 26, 2010.The<strong>Quinquennium</strong> Golden2007-2012Garima-IIShresth3
The<strong>Quinquennium</strong> Golden2007-2012Three cell lines of buffalo ES cells were produced, which areat 165, 125 and 115 passages and have been in culture over34, 26 and 24 months, respectively. These are the only buffaloES cell lines available in the world. These cell lines can now beused for transgenesis, cloning etc.Characterization of buffalo embryonic stem celllinesCloned transgenic buffalo embryos were produced usingbuffalo somatic cells and ES cells. ES cells were derived froman adult buffalo whose productive and reproductive featureswere well characterized. A two-round nuclear transfer (NT)approach was applied, wherein the first round NT providedthe cloned blastocysts from which NT-ES cell-like cells werederived and used as nuclear donor for a second round of NT.Buffalo adult fibroblasts and NT-ES cell-like cells of the samegenotype were transfected and were further used in Handguidedcloning for production of cloned transgenic embryos.Pregnancy established using oocytes collectedfrom live Sahiwal cattle by Ovum-Pick-upOocytes were collected from the ovaries of Sahiwal cattleby follicular aspiration using an ultrasound machine with atransvaginal convex transducer with a needle guide, single lumen18-gauge 55 cm long sterile needle with an ultrasound echotip and a vacuum pressure of 80 mmHg. Oocytes were thensubjected to in vitro maturation, in vitro fertilization and invitro culture for development of embryos. When the embryosdeveloped to transferable stage i.e., the blastocyst stage, thesewere transferred to the recipient i.e., the surrogate mother. Usingthis OPU and IVF technique, a pregnancy was established.A non-invasive DNA isolation methodology indairy animals developedThis is an innovative process for the isolation of GenomicDNA from fecal /epithelial tissue in cattle and buffalo. Theinvention relates to a process for purifying and/or isolatingnucleic acids from domestic animal feces (cattle and buffalo),Cloned transgenic blastocysts using transfected ES cell: A, GFP taggedembryonic stem cell colony (under light microscopy); b, Expression of GFP(under fluorescent microscopy; C, Blastocyst developed using hand guidedcloning and D, Expression of GFP in blastocyst (fluorescent microscopy)(Scale bar represents 50 μm).Agarose (1%) gel elctrophoresis of genomic DNA isolated from buffalo fecaltissue (7) Genomic DNA sample (3 µl of each) electrophoresed in a 1%agarose gel with Lambda marker (M), (S) Skin, (K) Kidney, (B) Brain,(L) Liver, (M) Muscle4
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NDRI Quinquennium Book COVER FINAL CURVE.cdr

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