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Amersham 600 rgb imager

Manufactured by GE Healthcare
Sourced in United States

The Amersham 600 RGB imager is a multipurpose imaging system designed for a variety of life science applications. It utilizes a charge-coupled device (CCD) sensor to capture high-resolution images of fluorescent, chemiluminescent, or colorimetric samples.

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2 protocols using amersham 600 rgb imager

1

High-Molecular-Weight DNA Extraction

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The molecular weight of the extracted DNA was determined by running 2 μL of each sample in 0.8% agarose gel, 1× TAE buffer (Thermo Fisher Scientific) for 4 h at 60 V. The DNA was stained with 1× SYBR safe (Invitrogen, Waltham, Massachusetts, USA). The molecular weight of the genomic DNA was estimated by comparison with a HMW DNA ladder (Lambda DNA/EcoRI+HindIII; Thermo Fisher Scientific). Digital images of the gels were generated in an Amersham 600 RGB imager (GE Healthcare, Chicago, Illinois, USA) using automatic collection parameters. The images of the gels were enhanced (contrast, homogenization, and background removal) in ImageJ version 1.52a (Rasband, 2018) prior to the digital analysis. The 1D gel electrophoresis image analysis software GelAnalyzer2010a (http://www.gelanalyzer.com) was used to create profiles of the distribution of DNA fragments (Fig. 2A, B) using the Lambda DNA/EcoRI+HindIII DNA ladder as a size reference.
For all species, HMW DNA was exclusively observed in the DNA samples extracted from cells homogenized using the LN2 grinding method. The DNA extracted using this treatment was observed as a tight, clear band over the 21.2‐kbp marker band, whereas DNA extracted from cells homogenized using the other treatments displayed substantial smearing and lacked a clear HMW DNA band, consistent with high DNA fragmentation (Fig. 2).
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2

Quantification of Protein Expression

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Triplicate lysates (in 8 M urea) from the above proteomic cell line comparison were mixed with NuPAGE loading buffer (Invitrogen) and loaded onto a 4–12% Bis-Tris (NuPAGE, Invitrogen). Gels were washed and proteins were transferred to nitrocellulose (iBlot 2, default 7 min method). Membranes were washed and stained with Ponceau S (in 5% acetic acid) for 10 min to measure total protein. Excess stain was removed by washing in water. Membranes were incubated in 5% fat-free milk in TBST for 30 min at room temperature to remove the Ponceau S stain and block the membrane. Membranes were incubated in primary antibody (anti-ALPK2 (Abcam) or anti-ENG (Cell Signaling Technologies)) overnight at 4 °C. After primary antibody incubation, membranes were washed in TBST and water prior to incubation with secondary antibodies (1 h, room temperature). Blots were developed (SuperSignal West Pico PLUS Chemiluminescent Substrate) and analyzed using an Amersham 600 RGB Imager (GE). Captured images were processed using ImageJ for quantitative comparison of total protein abundance across cell lines.23 Correlation analysis was done using R.16
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