Miller chromatin spreading

A. The Miller Chromatin Spreading Method Overview and Brief History... 

Some of the protocols presented here are very similar to methods previously published (Osheim and Beyer, 1989 Beyer etal, 1994). The 1989 work is a more general consideration of the Miller chromatin spreading method, which predates our recent refinements and does not present details of oocyte injections. The 1994... 

Successful Miller chromatin spreads depend on using low-ionic-strength water that has an alkaline pH between 8.5 and 9.0, which we call pH 9 water. We previously used deionized, glass-distilled water, but because of too much variability we now purchase HPLC water, which has a very low ionic composition (Mallinckrodt ChromAR HPLC water, VWR MK 679510). All solutions are made using this water. We adjust the pH of all solutions using standard pH 10 buffer standard for pH meters (Fisher, cat. SB116-500), which is a 0.05 M... 

Miller chromatin spreading method. We consider our efforts to be still developing, as we attempt to maximize the visualization of specific, active, mappable genes. One of our main goals at this time is to find a DNA sequence element that will ensure efficient Pol II termination so that the common problem of read-through transcription (as seen in Fig. 6) can be overcome. We currently are testing three different elements reported to have roles in transcription termination. 

Mougey, E O Reilly, M Osheim, Y. N Miller, O. L.. Jr.. Beyer, A. L., and Sollner-Webb, B. (1993). The terminal balls characteristic of eukaryotic rRNA transcription units in chromatin spreads are rRNA processing complexes. Genes Dev. 7, 1609-1619. 

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