By Carlos Lois (auth.), Shirley Pease, Carlos Lois (eds.)
Mammalian and Avian Transgenesis offers a suite of novel equipment for the construction of quite a lot of transgenic animals. The handbook focuses mostly on mice, but in addition comprises protocols for profitable transgenesis in rats, cows, pigs and birds.
The handbook presents distinct, step by step protocols masking all features of the construction of transgenic animals, together with using lentiviral vectors in gene move, intracytoplasmic sperm injection, nuclear move, huge insert transgenesis, conditional gene expression structures, using reporter genes in transgenesis and transgenesis in huge animals and birds. The textual content is supplemented by way of incredible colour images. whereas the point of interest is on newly tested recommendations, the basic equipment of transgenesis also are coated for these new to the sphere. therefore this guide is ideally fitted for these wishing to undertake new applied sciences in transgenesis.
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Extra info for Mammalian and Avian Transgenesis — New Approaches
2. Penetration of zona pellucida. The piezo-driven microinjection pipette has penetrated through the zona pellucida after applying a few piezo pulses; notice the drilled section of the zona pellucida is now inside the tip of the microinjection pipette tip Fig. 3. Advance the tip of the pipette. C. Kent Lloyd Fig. 4. Expel sperm head. The oolema is broken by a single piezo pulse, and the sperm head is injected into the cytoplasm Fig. 5. Withdrawal of pipette. The microinjection pipette is withdrawn gently from the oocyte, leaving behind the sperm head 2 Intracytoplasmic Sperm Injection (ICSI) in the Mouse 35 1.
If the injection pipette can be set at an angle of 15–20ı against the injection dish, then a straight injection pipette may be used successfully. If this is not possible, then an improvement may be to put a bend in the injection pipette, such that the angle of penetration of embryos is more horizontal. In this case, the elbow should be far away from the tip of the pipette where the mercury is located. If the elbow is in the region of the mercury, then it will affect the ﬂow control of the ﬂuid and reduce the mercury-enhanced piezo vibration.
2002). , strain related) or that loses motility (Mazur et al. K. Lloyd unpublished data; (Wakayama et al. K. Lloyd unpublished data; (Wakayama and Yanagimachi 1998; Kusakabe et al. 2001)), snap freezing, or evaporative drying, and is thus not conducive for use in either IVF or AI, has been used successfully for ICSI. This latter fact has, in part, been responsible for the recent popularity of ICSI as a procedure for mouse strain and colony management. K. Lloyd unpublished data; (Szczygiel et al.