Science of Synthesis

Georgiev, T.; Cazzamalli, S.; Neri, D.; Bassi, G., Science of Synthesis: DNA-Encoded Libraries, (2024) 1, 491.

Encoding is crucially important for the construction of DELs. High yields of encoding enable the efficient and high-fidelity sequencing of library members by high-throughput DNA sequencing (HTDS). There are multiple ways to construct single-stranded DELs (ssDELs), which rely on different encoding strategies.[‌1‌–‌7‌] The most common approach is known as “DNA-recorded synthesis”[‌24‌] and is outlined in ‌Scheme 5‌. The main premise is that each chemical transformation performed on the nascent library is “recorded” by the addition of a DNA fragment (encoding step). This procedure, alternated with split-and-pool techniques, can be performed iteratively to allow the formation of large combinatorial diversity. In the first step, chemical building blocks (A) are introduced to a universal oligonucleotide bearing an appropriate functional group (FG), which enables the chemical reaction. The encoding step is performed via enzymatic splint-assisted ligation. The consecutive steps of chemical transformation and encoding can be iterated to allow the construction of libraries with different designs containing two, three, or more building blocks.

‌Meeeee 8‌ Meeeeeeeeeee eee Meeeeeee ee e Meeeee-Meeeeeee MMM eee MMM-Meeeeeee Meeeeeeee

Me eeeeeeeeeeee ee eee eeeeeeeee eeeeee-eeeeeeee-eeeeeeee eeeeeeeee ee eeeeeeeee ee ‌Meeeee 8‌. Meeeeeeeeeeeeee eeeeeee eeeeeee eeeeeee eeeeeeee eeeeee (eee eeeeeee) eee eeeeeeeeeeee eeeeeee eeee eeeeee MMM-eeeeeeee (Meee M) ee eeeee ee ee eeeeeee (eeeeee). Mee eeeeeee eeee ee eeeeeeeeeee eeeeeeee ee eeeee ee eeeeeeeeeee eeeeee ee eeee eee eeeeeee eeeeeee eee eee Meee M eeeeeeee, eee ee eeeeee eeeeee ee eeee eee eeeeeee eeeee eeeeeee eeeeeeee ee eeee eeeeeeeeeee. Mee Meee M eeeeeeeee eeee ee eeeeeeeeeeeeee ee eee 8′ eee ee eeeee eee eeeeeeeee ee eee eeeeeeeee eeeee eeee eeee eee eeeeeee eeeeeee. Meeeeeeee ee eeeeeeee ee eeeeeee eee eeeeeee ee eeeeeee eeeeeee, eeeeeee, eee Meee M eee e eeeee eeee, eeeeeeee ee eeeeeeeeee eeeeeee ee eeee eeeeeeeeeee (‌Meeeee 8‌, eeee 8). Meeeee eeee eeee, eee 8′-eeeeeee eee ee eee eeeeeee eeeeeee eeeeeeee ee eeeeeee eeee eeeee eeeeeeeee ee eee 8′-eeeeeeeeeeeeee eee ee eee eeee. Meee, eeeeeeee ee M8 MMM eeeeee, MMM, eee eee eeeeee eeeeee eeeeeeeee eee eeeeeeeee eeeeeeee, eeeeeeeeee eee eeeeeeeee eee eeeeeeeee eeeeeeeeeeeee (MMM) eeeeeeee. Mee 8′-MM eeeeeeeee e eeeeeeeeeeee eeeeeeeeeeee, eeeee eeeeeee eee eee eeeeeeeeeeeeeee eeeeeeeee eee eeeeeeeee e eeeeeeee ee MMM. Mee eeeeeee eeeeeeeeeeeeeeee eee eeeeeeeee eeeeeeee eee eeeeeee-eeeee MMMM.

‌Meeeee 8‌ Meeeee-Meeeeeee-Meeeeeee Meeeeeee Meeeeeeee

Mee eeee ee Meeeeee ee ee.[‌88‌] eee eeee eeeeee ee ee eeeeeee ee eeeeeeeee eeeeeee ee eee eeeeeeeeeeee eeeeeee ee eee eeeeee-eeeeeeee-eeeeeeee eeeeeeee. Mee eeeeeeeeee, eeee eeee 8 ee eeeeeee eeeeeeeeeeee, eeeeeeeee eeeeeeee, eeeeeeeeeeee, eee eeeeeeeeeeeeeeee, eeee ee eeeeeeeee (‌Meeeee 8‌). Mee eeeeee ee eee eeeeeee (eeeeee MM-MMM) eee eeeeeeee eeeeee eeeeeeeeeee ee 8-eeeee-8-(eeeeeeeeee)eeeeeeeee eeeee ee eee eeeeeee eeeeeeee eee eeeeeeeee ee eee eeee ee eeeeeeee eeeeee ee e eeeeeeeee eeeeeee, eeeeeeeeeeee ee e eeeee ee 888 888 eeeeeee.[‌88‌] Me eeeeee ee eeeee eeee, ee eeee eeee, eee eeeeeeee eeeeee eeee eeeeeeee eeeee ee eeeee eeeeeeeee eeeeeeeeeee. Meee eeeee eeeeeeeee eeee eeeeeeeee eeee eeeeeeeeeee (88% ee eeeeee) eeee eeeeeeee ee eee eeeeeee eeeeeeeeeeee. Me eeeeeeeeeee ee ‌Meeeee 8‌, eee eeeeee eeeeeeee eeee eeee 8 eee eeeee eee eeeeeee eeee eeeeeeee eeeeee M eeeeeee eeeeee e eeeeeee eeee (Meeeee eeeee eeeeeeee) ee ee eeeeee (Meeeeeeeeee eeeee eeeeeeee). Meeee eeeeeeee eeee Meeee M, eee eeeeeeeee eeeeeeeee eeee eeeeee eee eeeeeeee eee eeeeeee-eeeee MMMM ee eeee eee eeeee MM-MMM.

‌Meeeee 8‌ Meee 8 ee eee MM-MMM Meeeeeeeeeee, Meeeeeee, eee Meeeeeeeeeee[‌88‌]

Meeeeeee ee eee eeeeeeee ee eeee 8 eee eee MMM ee eeee eeeeeeeeeeee ee eeeee ee ‌Meeeee 8‌. Meee (e) eeeee eeeeeeeee eeeeee eeee 8 eeeeeeee (eeeeeee MM = 88.8 MMe; eeeeeee eeee −8 ee −88); eeee (e) eeeeeeee eee eeeeeeee eeeeeeee ee eee eeee eeeeee (eeeeeee MM = 88.8 MMe; eeeeeee eeee −88 ee −88).

‌Meeeee 8‌ MM Meeeeeee ee Meeeeeee ee Meee 8 eee eee MM-MMM [eee-eeeeeeeeeeee eeeeeee (eeeee e/e = 888–8888)][‌88‌]

‌Meeeee 8‌ eeeee ee MMMM eeeeeeeeeeee (eeeeeeeeee ee λ = 888 ee) ee eee MMM: eeeeeeee M (8–88.8 eee) eeeeeeee eee eeeeee eeeeeeeeeeeeeee eee eeeeeeeee Meeee M, eeeee eeeeeeee M (88.8–88 eee) eeeeeeee eeeeeee eeee (eeee 8) eeeeeeee. ‌Meeeee 8‌, eeee (e) eeeee eeeeeeee ee eee eeeee MMM eeee ee MM/MM (MMM eeeeeeee eeee; eeeeeee eeee ee eeee ee 88.8 eMe; eeeeee 88-eee; e = eeeeeeee eeeeee); eee eeee (e) ee eeeeeeeeeeeeeee {8% eeeeeee eee; 8 = eeeeeeeee eeeeeeeeeeeeeee [88 eeee eeeee (ee)], 8 = eeee eeee 8 (88 ee), 8 = eeee eeee 8 (88 ee)}.[‌88‌]

‌Meeeee 8‌ MMMM Meeeeeeeeeee ee eee MM-MMM (Meee 8)[‌88‌]

‌Meeeee 8‌ Meeeeeee ee Meeee MM-MMM Meee ee MM/MM eee Meeeeeeeeeeeeee[‌88‌]

‌Meeeee 8‌ eeeee eee eeeeeeeee MM/MM eeeeeeeeeeeee (eeeeeeeeee ee λ = 888 ee): eeee (e) eee eeeee eeeeeee ee eee eeeeeee (eeeeee), eeeeeeeee eeeee, eee eee eeeee eeeeeee eeee (eeeeee MMMM eeeeeeeeeeee); eeee (e) eeeeeeee M (eeeee MMMM eeeeeeeeeeee; eee eeeeeee eee eeeeeeeee eeeee); eee eeee (e): eeeeeeee M (eeeee MMMM eeeeeeeeeeee; eee eeeeeee eeeee eeeeeee eeee).[‌88‌]

‌Meeeee 8‌ MM/MM Meeeeeeeeeeee ee eee MM-MMM[‌88‌]

Mee MMM-eeeeeeee eeeeeeee eeeee ee eeeeeeeee eeeeee eeeeeeee eee eeee eeee eeee ee eeeeeeeee eeee-eeeeeee eeeeeeeee eeee eeeee ee eeee eeee ee eeeeeeee eeeeee. Mee eeeeeee ee eee MM-MMM, e eeeee-eeeeeeee-eeeee eeMMM ee eeeeeee eeeeeeeeeee (‌Meeeee 8‌).[‌88‌] Mee eeeeeee eeeeeeeeeee eeeee eeee ee eeeeeeee eeeeee (MMe), eeee e eeeeeeeeee eeeeee eee ee eeeee eee eeee eee. M eeeeeeeee ee eee eeee eee eeeeee ee eeee eee ee MMe ee eeeee ee ‌Meeeee 8‌. Mee eeeeeeeee eeeeeee eeeeeeeee e eeeee ee 8 888 888 eeeeeeeeeee eeeeeee eee eeeeeeee eee eeeee eeeeee-eeeeeeee-eeeeeeee eeee ee eeeeeeeeee eeee eee MM-MMM. Mee eeeeeeee eeeeee-eeeeeeee eeeeeeee eeeeeeeee eee eee eeee eee eee MM-MMM eeeeeeeeeeee, eeee eeeeeeeeee eee eee eeeeee ee eeeeeeeeeee ee M8 MMM eeeeee eeee (eeeee ee eeee eeee eee eeee eeeeeee: 888 eeeee eee eeee eeeeeeee eeeeee 8.8) eee eee eee eeeeeeee eeee (88 e ee eeeeeee ee 8 e). Me eeeee ee eeeeeeeeeeee, eeee eee eeeeeeee eee eee eeeeeee eeee ee eeeeeee-eeeee MMMM ee ee eeeeeeeee eeeeeee ee eee eeee eeeeeeeee eee MM-MMM.

‌Meeeee 8‌ Meeeeeeee Meeeeeeeeeeeee ee eee Meeee Meeeeeeeeee MM-MMM[‌88‌]

Meeeeeeeeeeee eee ee-eeeeeee eeeeeeee ee eeeeeeeeeee eeeeeeee ee eee eeeeee-eeeeeeee eeeeeeee.[‌88‌] Meee eeeeeeee eeeeeeee eeeeeeee MMM–MMM eeeeeeee eeeee ee e eeeeee-eeeeeeeee eeeee–eeeeee eeeeeeeeeeeee (MeMMM eeeee eeeeeeee; ‌Meeeee 8‌). Mee eeeeee eeeeeeee eeeeeeeeeeeeeeee eeee ee eeeeee eeeeeeeeeeee ee eee 8′ eee, eeeee eee eeeeeee eeee e MMM eeeeee eeeeeee ee eeeee eeeeeeeeee eeeee ee eee 8′ eee. Mee eeeeeeeee eee eeeeeeeee eeeeeeee eeeeeeeeee eee eeeee ee eee eeeee eeee ee ‌Meeeee 8‌.

‌Meeeee 8‌ Me Meeeeeeeeee Meeeee-Meeeee Meeeeeeeeeeeeee Meeeeeee Meeeeeeee, Meeeeeeee e Meeee Meeeeeee eeeeeee e 8′-Meeee eee e 8′-Meeeee[‌88‌]

Meeeeeee ee eeeeee eeeeeeee, eeeeeeee eeeeeeee ee eeee eeeeeeeee, eee ee ee eeeeee ee eeeeee eee eeeeeeee eeee eee eeeeeeee eeeeeee (ee eeeee ee ee eeee eee eee eeeeeeeeeeeeeee eeeeeee/eeeeee). Meeeeeeeeeeee eeeeeeeee eee ee eeeeeee ee eeeeeeeeeeeeee eee, eeeeeeeeeeee, eee eeeeeee MMM ee eeeeeeeee eeee eee eeeeeeeeeee ee eeeeeee eeeeeeeeeeeee. Meeeeeee eeeeeeeee ee eeeeeeee eeeeeeee eeee eee eeeeeeeeee ee eeeee eeeeeeeeeeeeee eee eeeeeeeeeee eee ee eeeeeeeee ee eee-eeeeeee eeeeeee eeeeeeeee eeeeeee eeeeeeeeeee. Mee eeeeeeeee ee eeeeee eee eeeeeee ee eee MMM eeeeeeee (eeeeeeeeeeeeeee eeeee) eeeeee eee eeeeeeee eeeee ee eee eeeeeeee eeeeee eeeeee eeeeeee eeeeeeeeeeee. Meeeeeeee eeee eeeeeeee eeeeee eeeeee MMM eeeeeeeeeeee ee eeeeeee. Me eeeeee ee eeeee eeee eeeeee ee eeeeeeee eeeeeeee eee eeeee eeeeeeee ee eeeeeeeeeeee eeeeee-eeeeeeee eeeeeeeeee.[‌88‌]

M eeeeeeeeee eeeeeee eeeeeeeee eeeee ee eeeeeeeeeeeeeee eeeeeeeee eeeeeee eeee eee eeee ee Meeeeeeeeeeee eee ee-eeeeeee[‌88‌] ee eeeee ee ‌Meeeee 88‌. Meee eeee ee eeeee, eee eeeeeeee eeeeeeeee eeeeeeeee eeeeeeee eeeee eeeee eeee, eeeeee eeee, eee eeeeee eeeeeeeeee, eee eeeeeeeee eeeeeeee eeeee eeeeee-eeeeeeee eeeeee eeeeeee ee eeeeeee. Mee eeeee-eeee eeeeeee eeeeeeeeee eeeeeeee eeeee eeeeeeee ee eee eeeeeeee ee eeeeeeeee eeeeeeee. Mee eeeeee eeee eeeeeeeeee ee eeeeeeeeeeeeee eee eeeeeeeeeeeeeee (MMMM) eeeeeeee. Meeeeeee eeeeeeeee eee eeeeeeeee eeeeeee ee Meeeeee 8.8.8.8.

‌Meeeee 88‌ Meeeeeeeee eeeeeee Meeeeeeee Meeee ee Meeeeeeeeeeeeee Meeeeeeee[‌88‌]

Meeeeeeeee eeee eeeeeeeeeeeee eeee Mee[‌88‌] eeee eeeeeeeeee eeee eee Meeeeeee Meeeeeee Meeeeee.

Meeeee-eeeeeeee eeeeeeeee eee eeee ee eeeeeeeeeee eeeeeeeee MMM-eeeeeeeee eeeeeeeeee, ee eeeeeeeee ee eeeeee ee Meeeeee 8.8. M eeeeeeeee eeeeeeeeeeeeee ee eee eeeeeee ee eeeeeeee ee ‌Meeeee 88‌. Me eeeee, eeee eee eeeee-ee-eee eeeeeeeeeeeeeeee eeeeeee eeeeeeee eeeeeeee eeeeeeeee eeeee ee “eeeeeeeee” eee eee eeeeeeee eeeeeeee ee eeeeeee eeeeeee eeeeeeeee. Meeeeeeee, eeeee MMM-eeeeeeee eeeeeeee eeeeee eee eeeeeeee eeee eeee-eeeeeeee eeeeeee eeeeeeeeeee eeee e eeeeeeee eeeeeeeeee eeeee (MM). Meee eeeeeee eeeeee eee eeeeeeee eeeeeeee eeeeeeee ee eeeee eeeeeeeee ee eeeeeee e eeeeee eeeeeeeee eeeeeeee, eeeeeee ee ee eeeeeeeee eeeeeeee eeeeeeeeeeeeee.[‌88‌,‌88‌] Meee eeeeeee eee ee eeeeeeee ee eeeeeeee eeMMMe ee eeeeeee eeeeeee. Meeeeee eeeeeeeeeeeee eeee eeee eeee ee eeeeeeeee eee eeeeeeeeeee ee eee eeeeeeeee eeeeeeee ee Me eee ee-eeeeeee,[‌88‌] eee eeee eeeeeeee e eeeeeeeee eeeeeeee eeeeeeeeeeee (eee Meeeeee 8.8.8.8).

‌Meeeee 88‌ MMM-Meeeeeeee Meeeeeeee[‌88‌–‌88‌]

Meeeeeeeee eeeeeee, eeeeee-eeeeeeee MMM eeeeeee eee eeeeeeeeee eeee ee eeeeeeeeee eeeeeeee[‌8‌–‌8‌] eee ee eee eeeeeeee eeeeeeeee eeeeeeee ee eee-eeeeeee eeeeeee MMM eeeeeeeeeeeeee. Meeeeeee, Meee eee ee-eeeeeee eeee eeeeeeeee ee eeeeeeee eeee eeeeeee eee eeeeeeeeeeeeeee eeeeeee eeeeee- eee eeeeee-eeeeeeee MMMe ee eee eee ee e eeeeeeeeee eeeeeeee eeeeeeeee eeeee MMM eeeeeeeeeeee (‌Meeeee 88‌).[‌88‌] M 8-(8-eeeeeeeeee)eeeeeeeee (MMMM) eeeeee, eeee ee M-eeeeeeeeee M=M eeee, eee eeeee-eeeeeeeee ee e eeeeeeee eeeeeee ee eee eeeeeeeeeeeee eeeeeeeeeeeeeee eeeeee (888 ee, MM-M eeeee eeeeeeeeeee). Mee eeeeeeeeeee eeee eeee eeeeeeee eee eeeeeee eeeeeeeeee eeee eeee eee eeeeeeee eeeeeeeeee ee eeeeeeeeeee eee eeeeeee ee 888 ee (MM-M eeeee), eeeeeee ee eee eeeeeee eeeeeeeee ee eee eeeeeeeeeeee. Mee eeeeeeeee eeeee eeeeeeee eeeeeee eeeeeee ee eeee eeeeeeeeeee.

‌Meeeee 88‌ Mee Meeee ee Meeeee-Meeeeeee MMM Meeeeeeee Meeeeeeeee: eee Meeee, Meeeeeeeeeee, Meeeeeeee ee Meeeeeee Meee Me Mee Meeeeeee ee eee Meeeeeeeeeee; eee Meeee Meeeeee Meeeeeeeeee eee Meeee-eeeeeeeeee Meeeeeeee Meeeee[‌88‌]

Meeeeeeeeeee Meeeeeeee

Meeeee-Meeeeeee Meeeeeee eeee Meeee M eee MMM Meeeeeeeeeee (‌Meeeee 8‌):[‌88‌]

Me eee eeeeeeeeeeeeeee eeeeeeeeee (8 eeee eeee, eee) eeeeeeee eeeee eeeeeeee eeeeeeeeeeeeee eeee eeeeeeee eeeeee M eeee eeeee e 888 µM eeeeeeee ee eeeeee-eeeeeeeeeeeeeee (eeeee M; 8′-MMMMMMMMMMMMMMMMMMM-8′) ee eM Meeeeeeee eeeee (8 µM, 8.8 eeee eee eeee eeeeeeee), e 888 µM eeeeeeee ee eeeeeeeeeeeeee-eeee M (8′-MMMMMMMMMMMMMMMMMMMMMMMMMMMM-8′; 88 µM, 8 eeee eee eeee eeeeeeee), eM Meeeeeeee eeeee (8 µM), eee 88e M8 MMM eeeeee eeeeeeee eeeeee (888 eM Meee-MMe, 888 eM MeMe8, 888 eM eeeeeeeeeeeeee, 88 eM MMM; 8 µM; eM 8.8). Mee eeeeeeee eeee eeeeee eee 8 eee ee 88 °M, eee eeee eeeeee ee ee eeeeee M8 MMM eeeeee (8 µM, 888 eeeee/eM, 8.8 eeeee eeee eeeeeeee) eee eeeee. Mee eeeeeeee eee eeee ee ee eee 8 e. Meee eeeeeeeee eeeeeeee eee eeeeeeeeeee ee eee eeeeeeee ee 8 M eeeeeee eeeeee (8 µM; eM 8.8) eee ee eeeeeee ee 88 °M eee 8 eee. Mee eeeeeeee eeeeeeeee eeee eeeeeeee ee MM/MM eeeeee eeeeeee ee eeeeee eeeeeeee eeeeeee. Me MM eeeeeee ee eeeee eeeeeeee eeee eee eeeeeeee ee eeeeeeeee ee ‌Meeeee 8‌. Mee eeeeeee eeeeeeeeeee eeee eeeeee (eeeee eeeeee = 88.8 eM), eeeeeeeeeeee, eeeeeeeeeeee ee MeMM eeeeeeee, eeeeeeeeeee ee 8.8 M eeeeeeeeeeeeeeee eeeeeee (MMMM) eeeeee (eM 8) eee eeeeeeee ee 88 °M eeeeeee-eeeee MMMM (‌Meeeee 8‌). Mee eeee eeeeeee eeee eeeeee M (8.8 M MMMM) eee eeeeee M [MeMM/M8M (8:8), 8.8 M MMMM], eeee e eeeeeeee (% ee eeeeee M): 8% eee 8 eee, 8% ee 88% ee 88 eeee, 88% ee 88% ee 8 eeee, 88% ee 888% ee 8 eeee, 888% eee 8 eeee, eee e eeee eeee ee 8.88 eM/eee. Mee eee ee eeeeeee-eeeee MMMM eeeeeee eee eeeeee eeeeeeeeee ee eeeeeee eeee eeee eeeeeeee-eeee eeeeeeeee (e.e., eeeeeee, eeeeee, MMM, eee eeeeeeeee eeeee). Meee eeee eeeeeeeee eeeeeee ee eee eeeeee ee eeeeeee ee eeeeee eeeeeeeee eeeeeeeeee. Me eeee eeee, eee eeeeeee eeeeeeeeeee ee eee eeeeeeeee eee eeeeeeeeee ee ee 88.8 °M. Meee eee eeeeeeeeeee ee eeeee eee eeeee eeeee, ee eeeeee eee eeeeeee eee eeeeeeeee eeeee (eeeeeeee M) ee eeeee eeeeee eee eeeeeee eeee (eeeeeeee M). Mee eeeeeeeeee eeeeeeee eee eee eeee eeee 8 ee eeeeeeeeee ee eeeeeeeee ee eee eeee-eeeee eee eeeee-eeeee eeeeeee eeeeeee (eeee eeeeeee eee eeeeeeeee eeee ee eee eeeee eeeeeee eeee eee, eeeeeeeee, eee eeeeeeeeeee eeeeeeeeee eeeee). Meeee eeeeeeeeeeee, eee eeeeeeeeee eeeeeeee eee eeeeeeeeeeee eee ee eeeeeee eeeeeeee ee MM/MM ee eee eeeeeeee eeeeeee eeee eee ee eeeeeee eee (Meeeeee 8 eee 8).

MeMMM Meeee Meeeeeee (‌Meeeee 8‌):[‌88‌]

M eeeee eee eeeeeeeee eeeeeeeeeeeee Me(M) eeeeeeee eeeeee (eeeeee ee eeeeeeee eeeeeee eeeee, 8.8–8.8 ee),[‌88‌] eee eeee ee e eeeee eeeeee ee 88.8 µM ee eeeeeee eee eeeee eeeeeeee. Mee eeeeeee eee eeeeeeee eeee e eeeeeeee eeeeeeeeee 88 µM ee eeee eeeeeeeeeeeeeee, 8.8 eM eeee{[8-(8-eeeeeeeeeeeee)eeeeeee-8-ee]eeeeee}eeeee (MMMMM), eee 88 eM MeMe8 ee M8M/MMMM (88:88). Mee eeeeeee eee eeeeeeeee ee e eeeeeeeeeee (888 eee) eee 8 e ee 88 °M. Mee eeeeeeeeeee eee eeeeeee eee eeeeeeee ee eeeeee eeeeeee eee eeeeeeeeeeeeeee ee eeeeeee-eeeee MMMM [Meeeee MMMM Meeeeeee; e8888 Meeeeeeeee Meeeee, 8888 Meeeeeeeee Meeee Meeeeeee, eeeeeeee eeee ee MMeeeee MMM M88 eeeeee (8.8 µe; 8.8 ee × 88 ee)].

Meeeeeeeee 8-(8-Meeeeeeeee)eeeeeeeee/Meeeeee Meeeeeeeeeee ee Meeeeeeee MMM (‌Meeeee 88‌):[‌88‌]

Me eeeeeeee eeeee-eeeeeeeeeeee (eeeeeeeeeeeee), e eeeeeee (eeeee eeeeee 88 µM) ee eee eeeeeee-eeeeeeeeee eeeeeeeeeeeeeee (888 eeee) eee eee MMMM-eeeeeeeeee eeeeeeeeee (888 eeee) ee 8M MMM eeeeee eeee 8.8 M MeMe eee eeeeeeeeee eeeee e MMM MM-8888 Meeeeeeeeee eeeeeeeeeee (888 ee) ee 8 °M eee 8 eee. Meeee eeeeeeeeeee, eee eeeeeee eee eeeeeeeee ee MMMM/MM eeee eeeeeeee: 8% ee 88% M (8 ee 8 eee), 88% ee 88% M (8 ee 8 eee), 88% ee 88% M (8 ee 8.8 eee), 88% M (8.8 ee 8.8 eee), 88% ee 8% M (8.8 ee 8.8 eee), 8% M (8.8 ee 88 eee) (eeeee eeeeeee M: 888 eM (M8M)8MMMM eee 8 eM Me8M ee M8M; eeeeeee M: MeMM). Meeeeeeeee eee eeeee 88%.

Me eeeeeeee eeeee-eeeeeeeee (eeeeeeeeeeeeee), e eeeeeeee eeeeeeeeee eee eeeee-eeeeee eeeeeeeeeeeeeee eeeeeee (8 µM eeeee eeeeeeeeeeeee) ee M8M/MeMM (8:8) eee eeeeeeeeee eeeee 8-M eeeeeeee MM eeeee (888 ee) ee 88 °M eee 88 eee. Meeee eeeeeeeeeee, eee eeeeeee eee eeeeeeeee ee MMMM/MM (eeee eeeeeeee ee eeeee). Meeeeeeeee eee eeeee 88%.

References