Chromatin Immunoprecipitation from Drosophila Ovaries

Ovaries were dissected from NASP¹/ Df(3R), NASP²/Df(3R) or OrR females fattened for two days on wet yeast in Ephrussi Beadle Ringers (EBR). Stage 12 egg chambers were isolated, re-suspended in LB3 [63 (link)] and sonicated using a Bioruptor 300 (Diagenode) for five cycles of 30s on and 30s off at maximal power. Lysates were treated with RNase and Proteinase K and genomic DNA was isolated via phenol-chloroform extraction. qPCR was performed using primers previously described [64 (link)].

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Tirgar R., Davies J.P., Plate L, & Nordman J.T. (2023). The histone chaperone NASP maintains H3-H4 reservoirs in the early Drosophila embryo. PLOS Genetics, 19(3), e1010682.

Publication 2023

Bioruptor 300

Chloroform Females Genomic Ovaries Phenol Primers Proteinase k Rnase Yeast

Corresponding Organization : Vanderbilt University

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Variable analysis

independent variables

Genotype: NASP^1/Df(3R), NASP^2/Df(3R), or OrR females

dependent variables

Genomic DNA levels measured by qPCR

control variables

Egg chambers at stage 12
Ovaries dissected from females fattened for two days on wet yeast in Ephrussi Beadle Ringers (EBR)
Lysates treated with RNase and Proteinase K

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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