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[์—ฐ์„ธ๋Œฟ’ 10/10 A+] ๊ณตํ•™์ƒ๋ฌผํ•™๋ฐ์‹คํ—˜(2) 1์ฃผ์ฐจ PCR ๊ฒฐ๊ณผ๋ ˆํฌํŠธ

"[์—ฐ์„ธ๋Œฟ’ 10/10 A+] ๊ณตํ•™์ƒ๋ฌผํ•™๋ฐ์‹คํ—˜(2) 1์ฃผ์ฐจ PCR ๊ฒฐ๊ณผ๋ ˆํฌํŠธ"์— ๋Œ€ํ•œ ๋‚ด์šฉ์ž…๋‹ˆ๋‹ค.
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์ตœ์ดˆ๋“ฑ๋ก์ผ 2021.11.28 ์ตœ์ข…์ ฟ’์ž‘์ผ 2021.09
5P ๋ฏธ๋้ฉ๋ณด๊ธฐ
[์—ฐ์„ธ๋Œฟ’ 10/10 A+] ๊ณตํ•™์ƒ๋ฌผํ•™๋ฐ์‹คํ—˜(2) 1์ฃผ์ฐจ PCR ๊ฒฐ๊ณผ๋ ˆํฌํŠธ
  • ๋ฏธ๋้ฉ๋ณด๊ธฐ

    ์†Œ๊ฐœ

    "[์—ฐ์„ธ๋Œฟ’ 10/10 A+] ๊ณตํ•™์ƒ๋ฌผํ•™๋ฐ์‹คํ—˜(2) 1์ฃผ์ฐจ PCR ๊ฒฐ๊ณผ๋ ˆํฌํŠธ"์— ๋Œ€ํ•œ ๋‚ด์šฉ์ž…๋‹ˆ๋‹ค.

    ๋ชฉ์ฐจ

    1. Purpose
    2. Results
    3. Discussion
    4. Further Study

    ๋ณธ๋ๅฉ๋‚ด์šฉ

    Purpose:

    The purpose of the experiment is to amplify a small sample DNA sufficiently into a single PCR product to ensure all sizes of DNA can be amplified to completion through the same process.

    < ์ค‘ ๋žต >

    Figure 1 shows the DNA band of PCR sample after agarose gel electrophoresis along with two markers with given base pairs. The size of PCR products are conventionally calculated using the distance travelled on agarose gel after gel electrophoresis. Since only a digital copy of result image was given from the experiment, the tool Image Viewer on MATLAB was used to measure the distance travelled on gel in an arbitrary unit as shown in Figure 2. Figure 3 shows the calibration curve of distance travelled on agarose gel against the logarithmic value of DNA size in base pairs to the base of 10. The size of DNA fragments in base pairs is converted into logarithmic value to the base of 10 to ensure a linear graph is plotted instead of a polynomial graph.

    ์ฐธ๊ณ ์ž๋ฃŒ

    ยท Bessetti, Joseph. โ€œAn Introduction to PCR Inhibitorsโ€ Promega.Es, promega.es/-/media/files/resources/profiles-in-dna/1001/an-introduction-to-pcr-inhibitors.pdf?la=es-es. Accessed 12 Sept 2021.
    ยท Cha, Rita, and William Thilly. โ€œSpecificity, Efficiency, and Fidelity of PCRโ€. genome.cshlp.org, genome.cshlp.org/content/3/3/S18.full.pdf. Accessed 12 Sept 2021.
    ยท โ€œExperiment 3: Polymerase Chain Reaction (PCR)โ€. Southeastern, southeastern.edu/Academics/Faculty/jtemple/486/experiment%203.pdf. Accessed 11 Sept 2021.
    ยท "Gel Electrophoresis (Article) | Khan Academy". Khan Academy, khanacademy.org/science/ap-biology/gene-expression-and-regulation/biotechnology/a/gel-electrophoresis. Accessed 11 Sept 2021.
    ยท Kroemer, Tyasning. "How To Interpret DNA Gel Electrophoresis Results | Goldbio". Goldbio, goldbio.com/articles/article/Interpreting-Gel-Electrophoresis-Results. Accessed 12 Sept 2021.
    ยท "PCR Amplification | An Introduction To PCR Methods | Promega". Promega.Kr, promega.kr/resources/guides/nucleic-acid-analysis/pcr-amplification/. Accessed 11 Sept 2021.
    ยท Schrader, C. et al. "PCR Inhibitors - Occurrence, Properties And Removal". Journal Of Applied Microbiology, vol. 113, no. 5, July. 2012, pp. 1014-1026. Wiley, doi:10.1111/j.1365-2672.2012.05384.x. Accessed 13 Sept 2021.
    ยท "Top Ten Pitfalls In Quantitative Real-Time PCR Primer Probe Design And Use | Thermo Fisher Scientific - UK". Thermofisher, thermofisher.com/kr/ko/home/references/ambion-tech-support/rtpcr-analysis/general-articles/top-ten-pitfalls-in-quantitative-real-time-pcr-primer.html. Accessed 13 Sept 2021.
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      ํ•ดํ”ผ์บ ํผ์Šค FAQ ๋”๋ต–๊ธฐ

      ๊ผญ ์•Œ์•„์ฃผ์„ธ์š”

      • ์ž๋ฃŒ์˜ ์ •๋ณด ๋ฐ ๋‚ด์šฉ์˜ ์ง„์‹ค์„ฑ์— ๋Œ€ํ•˜์—ฌ ํ•ดํ”ผ์บ ํผ์Šค๋Š” ๋ณด์ฆํ•˜์ง€ ์•Š์œผ๋ฉฐ, ํ•ด๋‹น ์ •๋ณด ๋ฐ ๊ฒŒ์‹œ๋ฌผ ์ €์ž‘๊ถŒ๊ณผ ๊ธฐํƒ€ ๋ฒ•์  ์ฑ…์ž„์€ ์ž๋ฃŒ ๋“ฑ๋ก์ž์—๊ฒŒ ์žˆ์Šต๋‹ˆ๋‹ค.
        ์ž๋ฃŒ ๋ฐ ๊ฒŒ์‹œ๋ฌผ ๋‚ด์šฉ์˜ ๋ถˆ๋ฒ•์  ์ด์šฉ, ๋ฌด๋‹จ ์ „์žฌโˆ™๋ฐฐํฌ๋Š” ๊ธˆ์ง€๋˜์–ด ์žˆ์Šต๋‹ˆ๋‹ค.
        ์ €์ž‘๊ถŒ์นจํ•ด, ๋ช…์˜ˆํ›ผ์† ๋“ฑ ๋ถ„์Ÿ ์š”์†Œ ๋ฐœ๊ฒฌ ์‹œ ๊ณ ๊ฐ๋น„๋ฐ”์นด์ง€๋…ธ Viva์˜ ์ €์ž‘๊ถŒ์นจํ•ด ์‹ ๊ณ ๋น„๋ฐ”์นด์ง€๋…ธ Viva๋ฅผ ์ด์šฉํ•ด ์ฃผ์‹œ๊ธฐ ๋ฐ”๋ž๋‹ˆ๋‹ค.
      • ํ•ดํ”ผ์บ ํผ์Šค๋Š” ๊ตฌ๋งค์ž์™ฟ’ ํŒ๋งค์ž ๋ชจ๋‘๊ฐ€ ๋งŒ์กฑํ•˜๋Š” ์„œ๋น„์Šค๊ฐ€ ๋˜๋„๋ก ๋…ธ๋ ฅํ•˜๊ณ  ์žˆ์œผ๋ฉฐ, ์•„๋ž˜์˜ 4๊ฐ€์ง€ ์ž๋ฃŒํ™˜๋ถˆ ์กฐ๊ฑด์„ ๊ผญ ํ™•์ธํ•ด์ฃผ์‹œ๊ธฐ ๋ฐ”๋ž๋‹ˆ๋‹ค.
        ํŒŒ์ผ์˜ค๋ฅ˜ ์ค‘๋ณต์ž๋ฃŒ ์ €์ž‘๊ถŒ ์—†์Œ ์„ค๋ช…๊ณผ ์‹ค์ œ ๋‚ด์šฉ ๋ถˆ์ผ์น˜
        ํŒŒ์ผ์˜ ๋‹ค์šด๋กœ๋“œ๊ฐ€ ์ œ๋Œ€๋กœ ๋˜์ง€ ์•Š๊ฑฐ๋‚˜ ํŒŒ์ผํ˜•์‹์— ๋งž๋Š” ํ”„๋กœ๊ทธ๋žจ์œผ๋กœ ์ •์ƒ ์ž‘๋™ํ•˜์ง€ ์•Š๋Š” ๊ฒฝ์šฐ ๋‹ค๋ฅธ ์ž๋ฃŒ์™ฟ’ 70% ์ด์ƒ ๋‚ด์šฉ์ด ์ผ์น˜ํ•˜๋Š” ๊ฒฝ์šฐ (์ค‘๋ณต์ž„์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ๋Š” ๊ทผ๊ฑฐ ํ•„์š”ํ•จ) ์ธํ„ฐ๋„ท์˜ ๋‹ค๋ฅธ ์‚ฌ์ดํŠธ, ์—ฐ๊ตฌ๊ธฐ๊ด€, ํ•™๊ป“, ์„œ์  ๋“ฑ์˜ ์ž๋ฃŒ๋ฅผ ๋„์šฉํ•œ ๊ฒฝ์šฐ ์ž๋ฃŒ์˜ ์„ค๋ช…๊ณผ ์‹ค์ œ ์ž๋ฃŒ์˜ ๋‚ด์šฉ์ด ์ผ์น˜ํ•˜์ง€ ์•Š๋Š” ๊ฒฝ์šฐ
    ๋ฌธ์„œ ์ดˆ์•ˆ์„ ์ƒ์„ฑํ•ด์ฃผ๋Š” EasyAI
    ์•ˆ๋…•ํ•˜์„ธ์š”. ํ•ดํ”ผ์บ ํผ์Šค์˜ ๋ฐฉ๋Œ€ํ•œ ์ž๋ฃŒ ์ค‘์—์„œ ์„ ๋ณ„ํ•˜์—ฌ ๋‹น์‹ ๋งŒ์˜ ์ดˆ์•ˆ์„ ๋งŒ๋“ค์–ด์ฃผ๋Š” EasyAI ์ž…๋‹ˆ๋‹ค.
    ์ €๋Š” ์•„๋ž˜์™ฟ’ ๊ฐ™์ด ์ž‘์—…์„ ๋„์™ฟ’๋“œ๋ฆฝ๋‹ˆ๋‹ค.
    - ์ฃผ์ œ๋งŒ ์ž…๋ ฅํ•˜๋ฉด ๋ชฉ์ฐจ๋ถ€ํ„ฐ ๋ณธ๋ๅฉ๋‚ด์šฉ๊นŒ์ง€ ์ž๋™ ์ƒ์„ฑํ•ด ๋“œ๋ฆฝ๋‹ˆ๋‹ค.
    - ์žฅ๋ฌธ์˜ ์ฝ˜ํ…์ธ ๋ฅผ ์‰ฝ๊ณ  ๋น ๋ฅด๊ฒŒ ์ž‘์„ฑํ•ด ๋“œ๋ฆฝ๋‹ˆ๋‹ค.
    - ์Šคํ† ์–ด์—์„œ ๋ฌด๋ฃŒ ์บ์‹œ๋ฅผ ๊ณ„์ •๋ณ„๋กœ 1ํšŒ ๋ฐœ๊ธ‰ ๋ฐ›์„ ์ˆ˜ ์žˆ์Šต๋‹ˆ๋‹ค. ์ง€๊ธˆ ๋ฐ”๋กœ ์ฒดํ—˜ํ•ด ๋ณด์„ธ์š”!
    ์ด๋Ÿฐ ์ฃผ์ œ๋“ค์„ ์ž…๋ ฅํ•ด ๋ณด์„ธ์š”.
    - ์œ ์•„์—๊ฒŒ ์ ํ•ฉํ•œ ๋ฌธํ•™์ž‘ํ’ˆ์˜ ๊ธฐ์ค€๊ณผ ํŠน์„ฑ
    - ํ•œ๊ตญ์ธ์˜ ๊ฐ€์น˜๊ด€ ์ค‘์—์„œ ์ •์‹ ์  ๊ฐ€์น˜๊ด€์„ ์ด๋ฃจ๋Š” ๊ฒƒ๋“ค์„ ๋ฌธํ™”์  ๋ฌธ๋ฒ•์œผ๋กœ ์ •๋ฆฌํ•˜๊ณ , ํ˜„๋Œ€ํ•œ๊ตญ์‚ฌํšŒ์—์„œ ์ผ์–ด๋‚˜๋Š” ์‚ฌ๊ฑด๊ณผ ์‚ฌ๊ณ ๋ฅผ ๋น„๊ตํ•˜์—ฌ ์ž์‹ ์˜ ์˜๊ฒฌ์œผ๋กœ ๊ธฐ์ˆ ํ•˜์„ธ์š”
    - ์ž‘๋ณ„์ธ์‚ฌ ๋…ํ›„๊ฐ
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