Measurement Kit without processing in cultured cell DNA

DNA Quantity Kit


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Background

DNA can be quantified without purification process from cultured cell. Color Development Reagent binds to cellular DNA and fluoresces at 458 nm.

Kit component

Component Quantity Storage
Color Development Regent 10 mL 4ºC
Dilution Buffer 150 mL × 2 4ºC
DNA Standard (100 ug/ ml) 2 mL 4ºC

#Additional Materials Required
  EPurified water
  EFluorometer
  EPhosphate Buffered Saline (PBS)

Preparation of Standard Curve

1. Standard sample preparation
  Dilute the stock DNA Standard (100 ug/ ml) with purified water to 50, 25, 12.5 ug/ml.
  Use the purified water for a 0 ug/ ml and the DNA standard stock for 100 ug/ ml.
  Diluted sample standards can be stored frozen at -20°C.

2. Transfer each 50 ul of standard sample to new tubes.

3. Add 1ml Dilution Buffer to each standard sample and mix well.

4. Add 50 ul of Color Development Regent to each tube.

5. Mix thoroughly.

6. Measure fluorescence. (Excitation at 356nm, emission at 458nm)

Sample Analysis (24well plate format)

1. Remove culture medium and rinse the culture plate with Phosphate Buffered Saline (PBS).
2. Remove PBS and add 500 μl of dilution buffer to each well.
3. Sonicate cells until completely homogenated.
4. Transfer 50 ul of homogenate sample to new tube.
5. Add 1ml of Dilution Buffer to each tube and mix well.
6. Add 50 ul of Color Developer to each tube.
7. Mix thoroughly.
8. Measure fluorescence. (Excitation at 356nm, emission at 458nm)

References

> Kalgaonkar, S., Nishioka, H., Gross, H. B., Fujii, H. Keen, C. L., Hackman, R. M. Bioactivity of a Flavanol-rich Lychee Fruit Extract in Adipocytes and Its Effects on Oxidant Defense and Indices of Metabolic Syndrome in Animal Models. Phytother. Res. 24, 1223-1228 (2010)
> Okura, H., Komoda, H., Saga, A., Kakuta-Yamamoto, A., Hamada, Y., Fumimoto, Y., Lee, CM., Ichinose, A., Sawa, Y., Matsuyama, A. Properties of Hepatocyte-like Cell Clusters from Human Adipose Tissue-Derived Mesenchymal Stem Cells. Tissue Eng. Part C Methods. 16, 761-70 (2010)
> Saito,Y., Goto, M., Maya, K., Ogawa, N., Fujimori, K., Kurokawa, Y., Satomi, S. The Influence of Brain Death on Tissue Factor Expression in the Pancreatic Tissues and Isolated Islets in Rats. Transplant. Proc. 41, 307-310 (2009)

Product List

Product Name Cat# Quantity Price

DNA Quantity Kit DataSheet ,

PMC-AK06-COS

1 KIT ¥37,000
$494

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.



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