Abstract: An isolation assembly for use with coiled tubing is described. The isolation assembly has a check valve for providing selective fluid communication through from the coiled tubing, through the isolation assembly, and into a downhole tool, such a straddle packer. The isolation assembly includes a shuttle moveable within a housing, and plurality of ports to selectively provide fluid communication from within the isolation assembly below the check valve, through the ports, and into the annulus, thus allowing selective surface-controlled equalization of downhole equipment. Also described is a bottom hole assembly including the isolation assembly. An improved method of fracing a formation includes providing a check valve, thus improving the life of the coiled tubing and the safety of the operation, and reducing the time to perform a given downhole operation.

Abstract: The present invention provides data to demonstrate that the re-fusion, of a mammalian karyoplast to an enucleated in vivo ovulated oocyte, following an unsuccessful initial simultaneous electrical fusion and activation event offers an additional alternative and improvement in the creation of activated and fused nuclear transfer-capable embryos for the production of live offspring in various mammalian non-human species including goats, pigs, rodents, primates, rabbits and cattle. Additionally, multiple electrical pulses offers an alternative and more efficient activation method in a simultaneous fusion and activation methodology for viable offspring production in a animal nuclear transfer program.

Abstract: The present invention provides data to demonstrate that the fusion performance of a cell-line in procedures involving fusion and cleavage indices either alone or in combination are a means for selecting a cell lines that will be successful in a nuclear transfer or microinjection program. This technique and method of selecting a cell line offers an additional alternative and improvement in the creation of activated and fused nuclear transfer-capable embryos for the production of live offspring in various mammalian non-human species including goats, pigs, rodents, primates, rabbits and cattle.

Abstract: It has been found that caprine sperm factor, alone or in combination with adenophostin A, produces the Ca2+ oscillations characteristic of egg activation seen with fertilization. The introduction of caprine sperm factor and adenophostin A to eggs to produce reconstructed embryos has been optimized.

Abstract: An isolation assembly for use with coiled tubing is described. The isolation assembly has a check valve for providing selective fluid communication through from the coiled tubing, through the isolation assembly, and into a downhole tool, such a straddle packer. The isolation assembly includes a shuttle moveable within a housing, and plurality of ports to selectively provide fluid communication from within the isolation assembly below the check valve, through the ports, and into the annulus, thus allowing selective surface-controlled equalization of downhole equipment. Also described is a bottom hole assembly including the isolation assembly. An improved method of fracing a formation includes providing a check valve, thus improving the life of the coiled tubing and the safety of the operation, and reducing the time to perform a given downhole operation.

Abstract: The invention features a purified embryonic or fetal caprine somatic cell. The somatic cell can be in a preparation of embryonic or fetal caprine somatic cells. In addition, the cell can be used to derive an embryonic or fetal caprine somatic cell line. The somatic cell can be a genetically engineered caprine somatic cell, e.g., a somatic cell which includes a transgenic sequence, or which a nucleic acid has been introduced.

Abstract: The present invention provides data to demonstrate that the re-fusion, of a mammalian karyoplast to an enucleated in vivo ovulated cocyte, following an unsuccessful initial simultaneous electrical fusion and activation event offers an additional alternative and improvement in the creation of activated and fused nuclear transfer-capable embryos for the production of live off-spring in various mammalian non-human species including goats, pigs, rodents, primates, rabbits and cattle. Additionally, multiple electrical pulses offers an alternative and more efficient activation method in a simultaneous fusion and activation methodology for viable offspring production in a animal nuclear transfer program.

Abstract: The invention features methods of cryopreserving sperm. The methods include providing a sample of sperm; cooling the sperm to a first temperature; maintaining the sperm sample at the first temperature; cooling the sperm to a second temperature; maintaining the sperm at the second temperature; and storing the sperm at a third temperature. These methods can be used to cryopreserve sperm from mammals, e.g., transgenic mammals and can be to preserve sperm for subsequent artificial insemination or in vitro fertilization.

Abstract: The invention features cloned and transgenic mammals, e.g., goats, which are made by introducing the genome of a somatic cell into an enucleated oocyte, preferably a naturally matured oocyte which in telophase, to form a reconstructed embryo.

Abstract: The invention features methods of producing a mammal, e.g., a cloned or transgenic mammal. The method includes maintaining a reconstructed embryo in culture until the embryo is in the 2 to 8 cell stage of embryogenesis, transferring the embryo at the 2 to 8 cell stage into a recipient mammal, and allowing the embryo to develop into a mammal.

Abstract: The invention features methods of cryopreserving sperm. The methods include providing a sample of sperm; cooling the sperm to a first temperature; maintaining the sperm sample at the first temperature; cooling the sperm to a second temperature; maintaining the sperm at the second temperature; and storing the sperm at a third temperature. These methods can be used to cryopreserve sperm from mammals, e.g., transgenic mammals and can be to preserve sperm for subsequent artificial insemination or in vitro fertilization.