ExiProgen™

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▶ 실험 자료

1Protein expression and purification

• 사용한 DNA 농도: 6 - 10 μg (purity > 1.8)
• Protocol number : 902
• Total run time : < 6 h
• yield : <100 μg

실험 과정

SDS-PAGE analysis
	<동일 단백질(GFP) 16 well 동시 발현/정제> Figure 1. GFP protein (His-tagged protein)'s expression/purification with ExiProgen™ M; AccuLadder™ Protein Size Maker (Low), E; expression sample P; purification sample
	<다른 종류 단백질 동시 발현/정제> Figure 2. His-tagged proteins expression/purification with ExiProgen™ M; AccuLadder™ Protein Size Maker (Low) E; expression sample, P; purification sample

2Total RNA Extraction From Tissue or cultured mammalian cell

• Sample: 15 ㎎ (rat liver tissue) or 1 X 10⁶ cells (HeLa)
• Protocol number: 202
• Average yield: 25 ㎍ (rat liver tissue), 10 ㎍ (HeLa)
• Average RQS: 9.5 이상

※ RQS(RNA quality score): 추출된 RNA의 quality를 수치화하여 나타낸 값으로 각 rRNA 피크의 높이 및 넓이, 분해된 RNA fragment 피크 등을 종합적으로 계산하여 표시합니다. Agilent사에 의해 개발된 RIN(RNA integrity number)와 동일한 개념이며, 보통 0부터 10까지 수치로 나타나는데 10에 가까울수록 RNA의 quailty가 좋은 것으로 판단할 수 있습니다.

실험 과정

• Rat의 조직에서 추출된 total RNA의 분석 결과

Figure 3. rRNA의 각 subunit이 잘 관찰되는 것으로 보아 total RNA 추출 과정 중 RNA의 분해 없이 intact하게 RNA가 추출된 것을 확인할 수 있습니다.

• Rat의 조직에서 추출한 Total RNA를 사용하여 Real-Time PCR을 수행한 결과

Figure 4. AccuPower^® RocketScrpt ™ RT PreMix(K-2101, Bioneer)와 AccuPower^® DualStar™ qPCR PreMix(K-6110, Bioneer) 그리고 Exicycler™ 96 Real-Time Quantitative Thermal Block(A-2060)를 이용하여 real time RT-PCR을 수행한 결과입니다.

• HeLa cell로 부터 추출한 total RNA의 분석 결과

Figure 5. HeLa cell (1 X 10⁶ cells)로부터 total RNA를 추출한 후 LabChip GX (Caliper Life Science)를 이용하여 추출된 RNA의 quality를 분석한 결과입니다. rRNA의 각 subunit이 잘 관찰되는 것으로 보아 total RNA 추출 과정 중 RNA의 분해 없이 intact한 RNA가 추출되었음을 알 수 있습니다. 또한, 평균 수율은 10 ㎍이며 평균 RQS는 9.5이상으로 나타났습니다.

• HeLa cell로 부터 추출한 total RNA를 사용하여 real-time PCR을 수행한 결과

Figure 6. HeLa cell (1 X 10⁶ cells)로부터 total RNA를 추출한 후 AccuPower^® RocketScrpt ™ RT PreMix (K-2101, Bioneer)와 AccuPower^® DualStar ™ qPCR PreMix (K-6110, Bioneer) 그리고 Exicycler™ 96 Real-Time Quantitative Thermal Block (A-2060)를 이용하여 real-time RT-PCR을 수행한 결과입니다.

3Genomic DNA from cultured mammalian cell (HeLa)

• Sample volume: 1X10⁶ cells
• Protocol number: 107
• Total prep time: about 1hr 30min.
• Average yield: 4-8 μg

실험 과정

• Agarose gel analysis

Figure 7. Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 1X10⁶ cells of cultured HeLa cell and lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have similar yields. Purity was also tested and was consistently between 1.8 and 2.0 (not shown).

4Genomic DNA from bacteria (E.coli)

• Sample volume: 1X10⁹ cells
• Protocol number: 109
• Total prep time: about 1hr 40min.
• Average yield: 8-12 μg

실험 과정

• Agarose gel analysis

Figure 8. Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 1X10⁹ cells of E.coli cells and lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have similar yields. Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

5Genomic DNA from tissue (bovine skeletal muscle)

• Sample volume: ~25 mg
• Protocol number: 102
• Total prep time: about 1hr 30min.
• Average yield: Up to 20 µg

실험 과정

• Agarose gel analysis

Figure 9. Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 25 mg of bovine tissues and lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have similar yields. Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

6Genomic DNA from plant tissue (spinach leaf)

• Sample volume: ~ 100 mg
• Protocol number: 104
• Total prep time: about 1 hr
• Average yield: Up to 5 µg

실험 과정

• Agarose gel analysis

Figure 10. Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 100 mg of spinach leaves and lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH₂O as a negative control in DNA extraction. Note all samples have similar yields. Purity was also tested and was consistently between 1.9 and 2.0 (not shown).