๐Ÿงช Research Tool

Optimal Primer Design

์œ ์ „์ž ์ด๋ฆ„์„ ์ž…๋ ฅํ•˜๋ฉด off-target์ด ์ตœ์†Œํ™”๋œ ์ตœ์ ์˜ PCR primer๋ฅผ ์ž๋™์œผ๋กœ ์„ค๊ณ„ํ•ฉ๋‹ˆ๋‹ค.
v2.0: Nearest-Neighbor Tm + ฮ”G ๊ธฐ๋ฐ˜ ํ‰๊ฐ€ + Hard Filter/Soft Scoring ์ฒด๊ณ„ ์ ์šฉ

๐Ÿงฌ

Primer ์„ค๊ณ„ ์ค€๋น„ ์™„๋ฃŒ

์œ ์ „์ž๋ฅผ ์ž…๋ ฅํ•˜๊ณ  ์„ค๊ณ„ ๋ฒ„ํŠผ์„ ํด๋ฆญํ•˜์„ธ์š”.
์ตœ์ ์˜ primer pair๋ฅผ ์ž๋™์œผ๋กœ ์ฐพ์•„๋“œ๋ฆฝ๋‹ˆ๋‹ค.

๐Ÿ“‹ ํƒ€๊ฒŸ ์œ ์ „์ž ์ •๋ณด

๐Ÿ”ฌ ์„ค๊ณ„ ์•Œ๊ณ ๋ฆฌ์ฆ˜ v2.0

Nearest-Neighbor Thermodynamics + ฮ”G ๊ธฐ๋ฐ˜ ํ‰๊ฐ€ + Hard Filter โ†’ Soft Scoring ์ฒด๊ณ„

1

์„œ์—ด ์ˆ˜์ง‘

NCBI RefSeq API๋กœ NM_ transcript ์„œ์—ด ํš๋“

2

ํ›„๋ณด ์ƒ์„ฑ

Sliding window + NN Tm ๊ณ„์‚ฐ (Salt correction ์ ์šฉ)

3

ฮ”G ํ‰๊ฐ€

Hairpin/Self-dimer/Heterodimer ฮ”G ๋ฐ Tm ๊ณ„์‚ฐ

4

Hard Filter

3' dimer, ฮ”Tm>3ยฐC, Hairpin Tmโ‰ฅannealing ๋“ฑ ์ œ๊ฑฐ

5

Soft Scoring

Tm, GC, Length, 3' end, Amplicon ๋“ฑ ์ ์ˆ˜ํ™”

6

Top 5 ์ถ”์ฒœ

์ ์ˆ˜ ์ˆœ ์ •๋ ฌ + ์ค‘๋ณต ์ œ๊ฑฐ ํ›„ ์ตœ์ข… ์ถ”์ฒœ