MCP๋กœ ์—ฐ๊ฒฐ โ†’

๊ณ„์‚ฐ ์ž…๋ ฅ

๊ณต์‹

Show calculation steps (1)
  1. Total DNA Yield

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

    C = concentration in ng/uL from the formula above; yield in micrograms

๊ด‘๊ณ 

๊ฒฐ๊ณผ

DNA ๋†๋„
250
ng/ยตL
๋†๋„ 250 ยตg/mL
์ด ์ˆ˜์œจ (๋ถ€ํ”ผ ๊ธฐ์ค€) 12.5 ยตg

DNA ๋†๋„ ๊ณ„์‚ฐ๊ธฐ๋ž€?

์ด ๊ณ„์‚ฐ๊ธฐ๋Š” NanoDrop์ด๋‚˜ ํ๋ฒณ ๋ฐฉ์‹์˜ UV ๋ถ„๊ด‘๊ด‘๋„๊ณ„๋กœ ์ธก์ •ํ•œ 260 nm ํก๊ด‘๋„(A260) ๊ฐ’์„ ์ด์šฉํ•ด ์‹œ๋ฃŒ ์† ์ด์ค‘๊ฐ€๋‹ฅ DNA(dsDNA)์˜ ๋†๋„๋ฅผ ์ถ”์ •ํ•ฉ๋‹ˆ๋‹ค. ํ•ต์‚ฐ์€ 260 nm์—์„œ ์ž์™ธ์„ ์„ ๊ฐ•ํ•˜๊ฒŒ ํก์ˆ˜ํ•˜๋ฉฐ, ์ด ํก๊ด‘๋„๋Š” ๋†๋„์— ์ •๋น„๋ก€ํ•ฉ๋‹ˆ๋‹ค. ๊ทธ๋ž˜์„œ ๋ถ„์ž์ƒ๋ฌผํ•™ ์‹คํ—˜์‹ค์—์„œ DNA๋ฅผ ๋น ๋ฅด๊ฒŒ ์ •๋Ÿ‰ํ•˜๋Š” ํ‘œ์ค€ ๋ฐฉ๋ฒ•์œผ๋กœ ๋„๋ฆฌ ์“ฐ์ž…๋‹ˆ๋‹ค.

DNA ์‹œ๋ฃŒ ํ๋ฒณ์„ ํ†ต๊ณผํ•˜๋Š” ๋ถ„๊ด‘๊ด‘๋„๊ณ„ ๊ด‘์„ ๊ณผ 260์—์„œ์˜ ํก๊ด‘ ํ”ผํฌ
DNA๋Š” 260 nm์—์„œ ์ž์™ธ์„ ์„ ๊ฐ€์žฅ ๊ฐ•ํ•˜๊ฒŒ ํก์ˆ˜ํ•˜๋ฉฐ, ์ด๊ฒƒ์ด A260 ์ธก์ •์˜ ๊ธฐ๋ฐ˜์ž…๋‹ˆ๋‹ค.

์‚ฌ์šฉ ๋ฐฉ๋ฒ•

๋ถ„๊ด‘๊ด‘๋„๊ณ„ ํ™”๋ฉด์— ํ‘œ์‹œ๋œ A260 ํก๊ด‘๋„ ๊ฐ’์„ ์ž…๋ ฅํ•˜์„ธ์š”. ๊ทธ๋‹ค์Œ ํฌ์„ ๋ฐฐ์ˆ˜๋ฅผ ์ž…๋ ฅํ•ฉ๋‹ˆ๋‹ค. ์ธก์ • ์ „์— ์‹œ๋ฃŒ๋ฅผ 1:10์œผ๋กœ ํฌ์„ํ–ˆ๋‹ค๋ฉด ๋ฐฐ์ˆ˜๋Š” 10์ด๊ณ , ํฌ์„ํ•˜์ง€ ์•Š๊ณ  ์›์•ก ๊ทธ๋Œ€๋กœ ์ธก์ •ํ–ˆ๋‹ค๋ฉด 1์„ ์ž…๋ ฅํ•˜๋ฉด ๋ฉ๋‹ˆ๋‹ค. ์‹œ๋ฃŒ์˜ ์ด ๋ถ€ํ”ผ(ยตL)๋ฅผ ํ•จ๊ป˜ ์ž…๋ ฅํ•˜๋ฉด ์ „์ฒด DNA ์ˆ˜์œจ๊นŒ์ง€ ์ถ”์ •ํ•  ์ˆ˜ ์žˆ์Šต๋‹ˆ๋‹ค. ๊ณ„์‚ฐ ๊ฒฐ๊ณผ๋กœ๋Š” ng/ยตL ๋†๋„(์ˆ˜์น˜์ƒ ๋™์ผํ•œ ยตg/mL ํฌํ•จ)์™€ ์ด ์งˆ๋Ÿ‰์ด ๋‚˜์˜ต๋‹ˆ๋‹ค.

๊ณต์‹ ์„ค๋ช…

๊ณ„์‚ฐ์‹์€ ๋‹ค์Œ๊ณผ ๊ฐ™์Šต๋‹ˆ๋‹ค.

$$\text{๋†๋„ (ng/ยตL)} = A_{260} \times 50 \times \text{ํฌ์„ ๋ฐฐ์ˆ˜}$$

์—ฌ๊ธฐ์„œ 50์€ dsDNA์˜ ๋ณ€ํ™˜ ์ƒ์ˆ˜๋กœ, ๊ด‘๊ฒฝ๋กœ 1 cm์—์„œ A260์ด ์ •ํ™•ํžˆ 1.0์ธ ์‹œ๋ฃŒ๊ฐ€ ์•ฝ 50 ng/ยตL์˜ ์ด์ค‘๊ฐ€๋‹ฅ DNA๋ฅผ ํ•จ์œ ํ•œ๋‹ค๋Š” ์˜๋ฏธ์ž…๋‹ˆ๋‹ค. ๋‹จ์ผ๊ฐ€๋‹ฅ DNA(ssDNA)์—๋Š” 33, RNA์—๋Š” 40์„ ์‚ฌ์šฉํ•˜์ง€๋งŒ, ์ด ๊ณ„์‚ฐ๊ธฐ๋Š” dsDNA๋ฅผ ๊ธฐ์ค€์œผ๋กœ ํ•ฉ๋‹ˆ๋‹ค. ๋งˆ์ดํฌ๋กœ๊ทธ๋žจ ๋‹จ์œ„์˜ ์ด ์ˆ˜์œจ์€ ๋‹ค์Œ๊ณผ ๊ฐ™์ด ๊ตฌํ•ฉ๋‹ˆ๋‹ค.

$$\text{Yield}_{\mu g} = \frac{C \times V}{1000}$$
A260 ร— 50 ร— ํฌ์„ ๋ฐฐ์ˆ˜ = ๋†๋„(ng/ยตL)๋ฅผ ๋‚˜ํƒ€๋‚ธ ๋„์‹
๋†๋„ ๊ณต์‹์€ A260์— 50 ng/ยตL ์ƒ์ˆ˜์™€ ํฌ์„ ๋ฐฐ์ˆ˜๋ฅผ ๊ณฑํ•ฉ๋‹ˆ๋‹ค.

๊ณ„์‚ฐ ์˜ˆ์‹œ

์˜ˆ๋ฅผ ๋“ค์–ด NanoDrop์—์„œ 1:10์œผ๋กœ ํฌ์„ํ•œ ์‹œ๋ฃŒ์˜ A260์ด 0.5๋กœ ์ธก์ •๋˜์—ˆ๊ณ , ์ตœ์ข… ๋ถ€ํ”ผ๊ฐ€ 50 ยตL๋ผ๊ณ  ๊ฐ€์ •ํ•ด ๋ด…์‹œ๋‹ค. ๋†๋„๋Š” ๋‹ค์Œ๊ณผ ๊ฐ™์Šต๋‹ˆ๋‹ค.

$$0.5 \times 50 \times 10 = 250 \text{ ng/ยตL}$$

์ด ์ˆ˜์œจ์€ ๋‹ค์Œ๊ณผ ๊ฐ™์ด ๊ณ„์‚ฐ๋ฉ๋‹ˆ๋‹ค.

$$\frac{250 \times 50}{1000} = 12.5 \text{ ยตg}$$

์ž์ฃผ ๋ฌป๋Š” ์งˆ๋ฌธ

์™œ 50์ธ๊ฐ€์š”? ๊ด‘๊ฒฝ๋กœ 1 cm, 260 nm ์กฐ๊ฑด์—์„œ dsDNA์— ๋Œ€ํ•ด ์‹คํ—˜์ ์œผ๋กœ ํ™•๋ฆฝ๋œ ํก๊ด‘ ๊ณ„์ˆ˜์ด๊ธฐ ๋•Œ๋ฌธ์ž…๋‹ˆ๋‹ค.

ํฌ์„ํ•˜์ง€ ์•Š์•˜๋‹ค๋ฉด ํฌ์„ ๋ฐฐ์ˆ˜๋Š” ์–ผ๋งˆ์ธ๊ฐ€์š”? \(1\)์„ ์ž…๋ ฅํ•˜์„ธ์š”.

ng/ยตL์™€ ยตg/mL๋Š” ๊ฐ™์€ ๊ฐ’์ธ๊ฐ€์š”? ๋„ค, ๋‘ ๋‹จ์œ„๋Š” ์ˆ˜์น˜์ƒ ์™„์ „ํžˆ ๋™์ผํ•ฉ๋‹ˆ๋‹ค. ๋”ฐ๋ผ์„œ 250 ng/ยตL๋Š” 250 ยตg/mL์™€ ๊ฐ™์Šต๋‹ˆ๋‹ค.

์ตœ์ข… ์—…๋ฐ์ดํŠธ: