结构基因组学

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 ...

This method for the simplified purification of mouse DNA is based upon an article entitled �Simplified mammalian DNA isolation procedure� by Peter W. Laird from Nuclei Acids Research Vol. 19 No. 15 pa ...

General precautions to be followed during handling of any genomic DNA sample: Avoid Shearing of the DNA by limiting mixing to inversion and gentle shaking. Vortexing of samples is not required to perf ...

1. Cut 0.5 cm of tail and mark mouse ears or toes2. Incubate at 55°C O/N in:500 l Tail Extraction Buffer 10 l Proteinase K (10 mg/ml)12.5 l 20% SDS 3. Add 150 l 5 M NaCl Vortex 5 minMicrofuge 15 min ...

1. Cut 0.5 cm of tail and mark mouse ears or toes2. Incubate at 55°C O/N in:500 l Tail Extraction Buffer 10 l Proteinase K (10 mg/ml)12.5 l 20% SDS 3. Add 150 l 5 M NaCl Vortex 5 minMicrofuge 15 min ...

The DNEasy Kits From Qiagen work well for preparing DNA from toes and tails. Alternatively the following protocol may be used. Cut the last phalange of one toe into 0.5 ml of lysis buffer (100 mM Tris ...

Although all of the DNA in an eukaryotic cell replicates during the S-phase of cell cycle there is a significant difference in the actual time in S-phase when a given chromosomal segment replicates. M ...

1 For cells cultured in 2D about 1-2X107 S1 cells were growth in 100mm dish and were cross-linked by adding formaldehyde to final concentration of 1% and incubated in room temperature for 10 minutes. ...

CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL FOR YEASTThis protocol is derived from a paper by Miriam Braunstein and is based on work in the Allis lab. The procedure was written by Pam Meluh and upda ...

CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL FOR YEASTThis protocol is derived from a paper by Miriam Braunstein and is based on work in the Allis lab. The procedure was written by Pam Meluh and upda ...