基因组学

This is the method I use routinely. ProcedureRemove tissue or fetus and fix in fresh 4% paraformaldehye/PBS (pH 7.0-7.5) for 1 hour at 4oC. Rinse three times for thirty minutes each with rinse buffer ...

This is the method I use routinely. ProcedureRemove tissue or fetus and fix in fresh 4% paraformaldehye/PBS (pH 7.0-7.5) for 1 hour at 4oC. Rinse three times for thirty minutes each with rinse buffer ...

Day -3 (Saturday night/early Sunday morning)Thaw out a vial of ES cells (R1 W9.5) and plate on a 6 cm dish which has previously been gelatinised and feeders laid down. Day -1 (Monday)Prepare 10 6 cm d ...

Media UsedTo prepare 100 ml mediumDMEM80 mlFCS15 mlNon-essential amino acids (100x)1 mlPen/strep (5000 1U/ml 5000 ug/ml)1 mlL-Glutamine 200 mM1 mlNucleosides stock (100x)1 mlBME 0.1M0.2 mlDulbecco's M ...

1. Introduction. This is a brief outline of the steps necessary to obtain transgenic mice or transgenic rats. Simply put the investigator constructs a transgene with a promoter and a structural gene ( ...

This is a two step procedure utilizing a Qiagen Large-Construct Kit for the initial purification followed by either a chromatographic separation OR gel purification. Qiagen Prep of BAC DNAModified fro ...

Since most people have their own preferences with this task you are completely at your leisure to ignore this section. This procedure works for me. However be aware that highly purified DNA is importa ...

This protocol was developed by Jan Parker-Thornburg at The Ohio State Universtiy1)Run the DNA digest on an agarose gel: Make a 1% Seaplaque (or other very high quality) Low melt agarose gel using 1X T ...

This protocol was developed by Jan Parker-Thornburg at The Ohio State Universtiy1)Run the DNA digest on an agarose gel: Make a 1% Seaplaque (or other very high quality) Low melt agarose gel using 1X T ...

This protocol was developed by Jan Parker-Thornburg at The Ohio State Universtiy1)Run the DNA digest on an agarose gel: Make a 1% Seaplaque (or other very high quality) Low melt agarose gel using 1X T ...

This protocol was developed by Jan Parker-Thornburg at The Ohio State Universtiy1)Run the DNA digest on an agarose gel: Make a 1% Seaplaque (or other very high quality) Low melt agarose gel using 1X T ...

This protocol was developed by Jon Neumann at the University of CincinnatiEar clip the mouse. Take the tissue and put into a microcentrifuge tube. Clean off clippers carefully between animals. Digest ...

This protocol was developed by Jon Neumann at the University of CincinnatiEar clip the mouse. Take the tissue and put into a microcentrifuge tube. Clean off clippers carefully between animals. Digest ...

Determine the age of your mice. Mice will usually not breed if they are younger than 4 weeks old. Similarly mice who have been housed alone or in pairs (with the same sex) will usually not breed if th ...

The purity of the transgene DNA used for microinjection is critical for the successful production of transgenic founder mice. DNA impurities in the form of bacterial endotoxins or organic contaminants ...

Introduction. We hope that this information will be useful to investigators who are unfamiliar with mouse breeding or who are breeding transgenic or gene targeted mice for the first time. These sugges ...

Heat ear punch in 300 µl 10 mM NaOH/0.1 mM EDTA at 95°C for 10 min. Store at RT. (300 µl aliquots of 10 mM NaOH/0.1 mM EDTA can be frozen. However do not store 10 mM NaOH/0.1 mM EDTA at room temperatu ...

You must demonstrate that you can detect the transgene to ensure that 1) you can genotype the founder mice and 2) to ensure that you will be able to maintain the lines without loss.Our recommendation ...

PCR screens must be designed to detect transgene DNA at the single copy level. Southern Blots analysis of transgenic mice need copy standards to estimate copy number. Copy standards are prepared by mi ...

whole mount preparationsimage of a whole mount from a 4 wk old virgin ProtocolSpread tissue on glass slideFix in Carnoy's fixative for 2 to 4 hours at r.t.Wash in 70 % EtOH for 15 minChange gradually ...