MCP๋กœ ์—ฐ๊ฒฐ โ†’

๊ณ„์‚ฐ ์ž…๋ ฅ

๊ณต์‹

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  1. Total RNA Yield

    Total RNA Yield: RNA ๋†๋„ ๊ณ„์‚ฐ๊ธฐ

    Yield in micrograms = concentration (ng/uL) x sample volume (uL) / 1000.

๊ด‘๊ณ 

๊ฒฐ๊ณผ

RNA ๋†๋„
10
ng/ยตL
๋†๋„ (ยตg/mL) 10 ยตg/mL
์ด RNA ํšŒ์ˆ˜๋Ÿ‰ 1 ยตg

RNA ๋†๋„ ๊ณ„์‚ฐ๊ธฐ๋ž€?

์ด ๊ณ„์‚ฐ๊ธฐ๋Š” NanoDrop ๊ฐ™์€ UV ๋ถ„๊ด‘๊ด‘๋„๊ณ„๋กœ ์ธก์ •ํ•œ 260 nm ํก๊ด‘๋„(A260) ๊ฐ’์„ ๋ฐ”ํƒ•์œผ๋กœ RNA ์‹œ๋ฃŒ์˜ ๋†๋„๋ฅผ ์ถ”์ •ํ•ฉ๋‹ˆ๋‹ค. ๋‹จ์ผ๊ฐ€๋‹ฅ RNA์— ์ ์šฉ๋˜๋Š” ํ‘œ์ค€ ํก๊ด‘ ํ™˜์‚ฐ ๊ณ„์ˆ˜๋ฅผ ์‚ฌ์šฉํ•˜๋ฉฐ, A260๊ฐ€ 1.0์ผ ๋•Œ ์•ฝ 40 ng/ยตL์˜ RNA์— ํ•ด๋‹นํ•œ๋‹ค๋Š” ์›๋ฆฌ๋ฅผ ๋”ฐ๋ฆ…๋‹ˆ๋‹ค. ํŠน์ • ๊ตญ๊ฐ€์— ํ•œ์ •๋˜์ง€ ์•Š๋Š” ๋ฒ”์šฉ ๋„๊ตฌ๋กœ, ์ „ ์„ธ๊ณ„ ์–ด๋А ์‹คํ—˜์‹ค์—์„œ๋‚˜ ๊ทธ๋Œ€๋กœ ์‚ฌ์šฉํ•  ์ˆ˜ ์žˆ์Šต๋‹ˆ๋‹ค.

์‚ฌ์šฉ ๋ฐฉ๋ฒ•

์ธก์ •ํ•œ A260 ๊ฐ’๊ณผ ์ธก์ • ์ „์— ์ ์šฉํ•œ ํฌ์„ ๋ฐฐ์ˆ˜๋ฅผ ์ž…๋ ฅํ•˜์„ธ์š”(ํฌ์„ ์—†์ด ์›์•ก ๊ทธ๋Œ€๋กœ ์ธก์ •ํ–ˆ๋‹ค๋ฉด 1์„ ์ž…๋ ฅํ•ฉ๋‹ˆ๋‹ค). ์‹œ๋ฃŒ ๋ถ€ํ”ผ(ยตL)๋ฅผ ์ถ”๊ฐ€๋กœ ์ž…๋ ฅํ•˜๋ฉด ์ด RNA ํšŒ์ˆ˜๋Ÿ‰๊นŒ์ง€ ๊ณ„์‚ฐ๋ฉ๋‹ˆ๋‹ค. ๊ณ„์‚ฐ๊ธฐ๋Š” ng/ยตL ๋‹จ์œ„์˜ ๋†๋„(ยตg/mL์™€ ๋™์ผ)์™€ ํšŒ์ˆ˜๋œ RNA ์ด๋Ÿ‰์„ ํ•จ๊ป˜ ๋ณด์—ฌ์ค๋‹ˆ๋‹ค.

๊ณ„์‚ฐ ๊ณต์‹ ์„ค๋ช…

RNA ๋†๋„(ng/ยตL):

$$\text{Concentration} = \text{A260} \times 40 \times \text{Dilution} \;\;\text{(ng/}\mu\text{L)}$$

์—ฌ๊ธฐ์„œ ๊ณ„์ˆ˜ 40์€ 1 cm ๊ด‘๊ฒฝ๋กœ ์…€์—์„œ 260 nm ํŒŒ์žฅ์— ๋Œ€ํ•œ ๋‹จ์ผ๊ฐ€๋‹ฅ RNA์˜ ๊ฒฝํ—˜์  ํก๊ด‘ ๊ณ„์ˆ˜์ž…๋‹ˆ๋‹ค. ์ด ํšŒ์ˆ˜๋Ÿ‰(ยตg)์€ ๋‹ค์Œ๊ณผ ๊ฐ™์ด ๊ตฌํ•ฉ๋‹ˆ๋‹ค.

$$\text{Yield (}\mu\text{g)} = \frac{\left(\text{A260} \times 40 \times \text{Dilution}\right) \times \text{Volume }(\mu\text{L})}{1000}$$
Diagram showing UV light passing through an RNA sample cuvette to a detector, with A260 absorbance multiplied by 40 and dilution factor
RNA concentration is derived from A260 absorbance using the 40 ng/ยตL factor and the dilution factor.

๊ณ„์‚ฐ ์˜ˆ์‹œ

A260 = 0.25, ํฌ์„ ๋ฐฐ์ˆ˜ = 50, ๋ถ€ํ”ผ = 100 ยตL์ธ ๊ฒฝ์šฐ๋ฅผ ๊ฐ€์ •ํ•ด ๋ด…์‹œ๋‹ค. ๋†๋„ \(= 0.25 \times 40 \times 50 = 500\) ng/ยตL. ์ด ํšŒ์ˆ˜๋Ÿ‰ \(= 500 \times 100 \div 1000 = 50\) ยตg์˜ RNA์ž…๋‹ˆ๋‹ค.

Bar comparison of pure RNA at A260/A280 ratio 2.0 versus contaminated sample with lower ratio
An A260/A280 ratio near 2.0 indicates pure RNA, helping validate the calculated concentration.

์ž์ฃผ ๋ฌป๋Š” ์งˆ๋ฌธ

์™œ 50์ด ์•„๋‹ˆ๋ผ 40์ธ๊ฐ€์š”? ๊ณ„์ˆ˜ 50์€ ์ด์ค‘๊ฐ€๋‹ฅ DNA์— ์‚ฌ์šฉ๋ฉ๋‹ˆ๋‹ค. ๋‹จ์ผ๊ฐ€๋‹ฅ RNA๋Š” ํก๊ด‘ ํŠน์„ฑ์ด ์กฐ๊ธˆ ๋‹ฌ๋ผ์„œ A260 ๋‹จ์œ„๋‹น 40 ng/ยตL๊ฐ€ ๊ณต์ธ๋œ ์ƒ์ˆ˜๋กœ ์“ฐ์ž…๋‹ˆ๋‹ค.

์ˆœ๋„๋Š” ์–ด๋–ป๊ฒŒ ํ™•์ธํ•˜๋‚˜์š”? A260๋งŒ์œผ๋กœ๋Š” ๋†๋„๋งŒ ์•Œ ์ˆ˜ ์žˆ์Šต๋‹ˆ๋‹ค. ์ˆœ์ˆ˜ํ•œ RNA์˜ ๊ฒฝ์šฐ A260/A280 ๋น„์œจ์ด ์•ฝ 2.0์ด๋ฉฐ, A260/A230 ๋น„์œจ๊ณผ ํ•จ๊ป˜ ํ™•์ธํ•˜๋ฉด ์˜ค์—ผ ์—ฌ๋ถ€๋ฅผ ํŒ๋‹จํ•  ์ˆ˜ ์žˆ์Šต๋‹ˆ๋‹ค.

๊ด‘๊ฒฝ๋กœ ๊ธธ์ด๋„ ์˜ํ–ฅ์„ ์ฃผ๋‚˜์š”? ์ด ๊ณ„์ˆ˜๋Š” ํ‘œ์ค€ 1 cm ๊ด‘๊ฒฝ๋กœ๋ฅผ ์ „์ œ๋กœ ํ•ฉ๋‹ˆ๋‹ค. ์ตœ์‹  ๋งˆ์ดํฌ๋กœ๋ณผ๋ฅจ ์žฅ๋น„๋Š” ์ธก์ •๊ฐ’์„ ์ž๋™์œผ๋กœ 1 cm ๊ธฐ์ค€์œผ๋กœ ๋ณด์ •ํ•ด ์ค๋‹ˆ๋‹ค.

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