By Jill C. Sible, Brian N. Wroble (auth.), David J. Carroll (eds.)
As the variety of sequenced genomes keeps to extend, knowing the services of newly came across molecules would require better potency and additional examine in the context of reside cells. In Microinjection: equipment and Applications, professional researchers give a contribution tools using microinjection options starting from expression of RNA to the combination of DNA into the genome with the final word aim of studying approximately gene expression, sign transduction, and protein functionality inside those residing cells. this flexible quantity updates confirmed innovations corresponding to cRNA expression in Xenopus oocytes, and examines new, state of the art applied sciences, together with antisense morpholino oligonucleotides, RNAi for knockdown experiments, and using integrase to supply transgenic animals, throughout microinjection thoughts that may be simply followed through any lab. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, chapters contain short introductions to the themes, lists of the mandatory fabrics and reagents, step by step, with no trouble reproducible laboratory protocols, and Notes sections, highlighting pointers on troubleshooting and fending off recognized pitfalls.
Comprehensive and simply available, Microinjection: equipment and Applications is a perfect resource for brand spanking new principles and strategies bound to relief within the improve of organic research.
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Additional info for Microinjection: Methods and Applications
To improve yields and efficiency of recovery even further, set up duplicate or triplicate reactions, pool prior to phenol/chloroform extraction, and complete the procedure, adjusting volumes accordingly. In practice, yields are greater than 2–3 times that of a single reaction, probably due to more efficient recovery in the extraction and precipitation steps. 4. Prolonged exposure to cysteine will damage the eggs. 5. Some investigators find it useful to calibrate several needles ahead of time, but the calibration should be rechecked just 14 Sible and Wroble prior to injecting the embryos.
119, 45–54. Nieuwkoop, P. , Faber, J. (1967) Normal Table of Xenopus laevis, North-Holland Publishing Company, Amsterdam. Moon, R. , Christian, J. L. (1989) Microinjection and expression of synthetic mRNAs in Xenopus embryos. Technique – J. Methods Cell. Mol. Biol.. 2, 76–89. Sive, H. , Grainger, R. , Harland, R. M. (2000) Early Development of Xenopus laevis: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Heasman, J. (2002) Morpholino oligos: making sense of antisense?
1988) ‘Chemiluminescence’ Ellis Horwood Series on Biomedicine. 13. Sambrook, J. W. (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. 14. Campbell, K. and Swann, K. (2006) Ca2+ oscillations stimulate an ATP increase during fertilization. Devel. Biol. 298, 225–53. 15. , Whittingham, D. J. (1994) Spatiotemporal dynamics of intracellular [Ca2+]i oscillations during growth and meiotic maturation of mouse oocytes. Development 120, 3507–17. Chapter 3 Analysis of 14-3-3 Family Member Function in Xenopus Embryos by Microinjection of Antisense Morpholino Oligos Jeffrey M.
Microinjection: Methods and Applications by Jill C. Sible, Brian N. Wroble (auth.), David J. Carroll (eds.)