J. Inst. Electrostat. Jpn.

, 30, 1 (2006) 26 31

⺰   ᢥ㔚⇇ࠍ↪޿ߚ੃ൻᴺߩ㐿⊒ߣߘߩᔕ↪
ਛ㊁㆏ᒾ㧖㧘࿡⼱ᓀ㧖㧘㜞ፉ๺ೣ㧖㧘᩵ㅴม㧖,㧖㧖㧘᳓㊁ᓆ㧖,1㧔2005 ᐕ 8 ᦬ 1 ᣣฃઃ㧧2006 ᐕ 1 ᦬ 17 ᣣฃℂ㧕Electrostatic Emulsification by AC Electric Field and BiologicalApplication
Michihiko NAKANO*, Sayaka KUNITANI*, Kazunori TAKASHIMA*,Shinji KATSURA*,** and Akira MIZUNO*,1(Received August 1, 2005; Accepted January 17, 2006)

Water-in-oil (W/O) emulsion has been used for biological and combinatorial application. In these applications, manywater droplets are served as micro reactors and several reactions are carried out simultaneously. To make W/Oemulsion, stirring device or homogenizer are used conventionally. Since these devices such as magnetic stirrer bar
are not disposable, they have many risks of contamination. In addition, these methods require a certain amount ofvolume at least 1 mL. In this paper, it was investigated that novel emulsification methods using AC voltage. There
were two approaches for emulsification: electrodes placed inside and outside of micro-tube. For the internal
electrode method, high voltage electrode was placed in micro-tube. On the other hand, for the external electrodemethod, high voltage and ground electrodes were placed on outside wall of micro-tube. In the both cases, when AC
voltage was applied to the electrode, water solution that was placed at bottom of the micro-tube was dispersed into oilphase. Moreover, it was confirmed that this emulsification did not affect polymerase chain reaction. Thesemethods have some features: preparing electrode inexpensively, especially for external method, non-contact operation
to prevent contamination from mixing devices and requiring small amount of volume, for example, 100 2L to 500 2L.
1.

ߪߓ߼ߦࠍหᤨߦ෻ᔕߐߖࠆ㧘ࠦࡦࡆ࠽࠻࡝ࠕ࡞ࠤࡒࠬ࠻࡝࡯ߦᆭജ

W/O㧔water-in-oil㧕ࠛࡑ࡞࡚ࠫࡦߪ㧘ᓸዊߥ᳓Ṣ߇ᴤਛߦࠍ⊒ើߔࠆ 1)㧚߹ߚ㧘ᓸዊߥⓨ㑆ߢ෻ᔕࠍⴕ߁ߎߣߢ㧘ߘߩಽᢔߒߡ޿ࠆ⁁ᘒߢ޽ࠆ㧚ㄭᐕ㧘ߎߩ W/O ࠛࡑ࡞࡚ࠫࡦࠍౝㇱߩၮ⾰Ớᐲߪ╬ଔ⊛ߦ㜞Ớᐲߦߥࠅ㧘ߘߩਛߢ⿠ߎࠆൻ

೑↪ߒߚ෻ᔕ♽߇ᵈ⋡ࠍᶎ߮ߡ޿ࠆ㧚ߎࠇߪ㧘ᓸዊߥ᳓Ṣߩቇ෻ᔕ߇㜞ല₸ൻߐࠇࠆ㧚ߎࠇߩ૞↪ࠍ೑↪ߒߚᣇᴺߣߒߡ㧘߭ߣߟ߭ߣߟࠍࡑࠗࠢࡠ࡝ࠕࠢ࠲࡯ߣߒߡ೑↪ߒ㧘ߘࠇߙࠇ

1 ಽሶ PCR㧔polymerase chain reaction㧕ᴺ߇᜼ߍࠄࠇࠆ 2)㧚

ߩ᳓Ṣߦ೎‫⹜ߩޘ‬ᢱࠍ౉ࠇߡ෻ᔕࠍⴕ߁ᣇᴺߢ޽ࠆ㧚W/O ࠛߎߩࠃ߁ߥ W/O ࠛࡑ࡞࡚ࠫࡦࠍᔕ↪ߒߚ෻ᔕ♽ߢߪ㧘ਥࡑ࡞࡚ࠫࡦࠍ↪޿ࠆߎߣߢ㧘࠙ࠚ࡞ࡊ࡟࡯࠻ࠍ↪޿ࠆࠃࠅ߽

ߦ᳓ṁᶧߣᴤ⋧ߣࠍࡑࠣࡀ࠹ࠖ࠶ࠢࠬ࠲࡯࡜࡯߿ࡎࡕࠫ࠽

◲නߦήᢙߩࡑࠗࠢࡠ࡝ࠕࠢ࠲࡯ࠍ⺞⵾ߢ߈ࠆ㧚଀߃߫㧘ㆮࠗࠩ࡯ࠍ૶ߞߡ੃ൻߐߖߡ޿ࠆ㧚ߎࠇࠄߩⵝ⟎ࠍ↪޿ࠆ႐ว㧘વሶ࡜ࠗࡉ࡜࡝࡯߆ࠄ㉂⚛ߥߤࠍ⋥ធ⊛ㅴൻ㧔directedߘߩᄢ߈ߐ߆ࠄ㧘޽ࠆ⒟ᐲߩኈ㊂㧔ዋߥߊߣ߽ 1mL㧕߇ߥ޿evolution㧕ߦࠃߞߡㆬᛯߔࠆߣ޿ߞߚࠃ߁ߥ㧘ᄙ⒳㘃ߩၮ⾰ߣ੃ൻߔࠆߎߣ߇ߢ߈ߕ㧘߹ߚ㧘ߘࠇࠄߩᯏ᧚ߪ➅ࠅ㄰ߒ૶↪ߐࠇࠆߎߣ߇ᄙ޿ߚ߼㧘․ߦ↢‛ቇ⊛⹜ᢱࠍ↪޿ࠆߣ߈ߦࠠ࡯ࡢ࡯࠼㧦੃ൻ㧘㔚᳇੃ൻ㧘ᶧṢ㧘ᓸ☸ൻ* ⼾ᯅᛛⴚ⑼ቇᄢቇࠛࠦࡠࠫ࡯Ꮏቇ♽㧔441-8580 ᗲ⍮⋵⼾ᯅᏒᄤષ↸㔕㓴ࡩਐ 1-1㧕Department of Ecological Engineering, Toyohashi University ofTechnology, 1-1 Hibarigaoka, Tempaku, Toyohashi, Aichi
441-8580, Japan.
** ᧲੩ᄢቇ㔚᳇Ꮏቇ⑼㧔113-8656 ᧲੩ㇺᢥ੩඙ᧄㇹ 7-3-1㧕
Department of Electrical Engineering, The University of Tokyo,
7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
1
mizuno@eco.tut.ac.jpߪ㧘ࠢࡠࠬ࡮ࠦࡦ࠲ࡒࡀ࡯࡚ࠪࡦߩේ࿃ߣߥࠆߣ޿ߞߚ໧㗴߇޽ࠆ㧚ߎߎߢߪ㧘ᯏ᪾⊛ߥᡬᜈⵝ⟎ࠍ૶ࠊߥ޿ᣇᴺߣߒߡ㧘㔚⇇ࠍ↪޿ߚ੃ൻߦߟ޿ߡᬌ⸛ߒߚ㧚ዋ㊂㧘଀߃߫ో㊂ 100㨪
5002L㧔᳓ṁᶧߪ 5㨪202L㧕ߢ੃ൻߔࠆߎߣ㧘ᡬᜈㇱಽ߇૶޿ᝥߡߦߢ߈ࠆߋࠄ޿቟ଔߢ޽ࠆߎߣ㧘޽ࠆ޿ߪ⹜ᢱṁᶧߣᡬᜈㇱಽ߇৻ಾធ⸅ߔࠆߎߣ߇ߥ޿㧘ߣ޿߁ࠃ߁ߥᡬᜈᣇᴺ27

࡞ࡒ࠾࠙ࡓ࠹࡯ࡊࠍᏎ޿ߡ㔚ᭂߣߒߚ㧚ߩᬌ⸛ࠍⴕߞߚ㧚㔚⇇ߣ੃ൻߣߩ㑐ଥߦߟ޿ߡ㧘㔚⇇ࠍ↪޿ߚ⸃੃ൻߪᄙߊ

2.1ታ㛎ᣇᴺߩ⎇ⓥ߇ⴕࠊࠇߡ޿ࠆ 㧚߹ߚ㧘㔚⇇ߦࠃࠆ੃ൻߦߟ޿ߡߪ㧘2.1.1

ࡁ࠭࡞వ┵ߦ㔚࿶ࠍශടߒߡㅪ⛯⊛ߦ⚦߆ߥ᳓Ṣࠍᴤ⋧ߦߎߎߢߪ㧘ᴤ⋧ߦ⇇㕙ᵴᕈ೷ࠍട߃ߡ⇇㕙ᒛജࠍᄌൻߐߖ3)4)ౝㇱ㔚ᭂᒻᚑߔࠆ㔚᳇੃ൻᴺ ߇޽ࠆ㧚ߒ߆ߒ㧘ߎߎߢ␜ߔࠃ߁ߥᴤ

ߚߣ߈ߩᲧセ߿㧘᳓⋧ߦ⚐᳓ߣ KCl ᳓ṁᶧࠍ↪޿ߡዉ㔚₸ߩ⋧ߣ᳓⋧ߣࠍ㕒ᱛߒߚ⁁ᘒ߆ࠄ੃ൻߔࠆᣇᴺߦߟ޿ߡߪ߶㆑޿ߦࠃࠆᲧセࠍⴕߞߚ㧚ߣࠎߤ⎇ⓥߐࠇߡ޿ߥ޿㧚ᧄ⺰ᢥߢߪ㧘㔚⇇ߦࠃߞߡ㕒⟎ߒࡊ࡜ࠬ࠴࠶ࠢ⵾ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߦߪ㧘1.5mL ࡑࠗࠢࡠ࠴ߚ⁁ᘒߩᴤ⋧ߣ᳓⋧ߣࠍᷙว࡮੃ൻߢ߈ࠆߎߣࠍ␜ߒ㧘ߘߩࡘ࡯ࡉ㧔ࠕࠗࡆࠬ㧕ࠍ↪޿㧘⹜ᢱߩ㊂Ყߪᴤ⋧㧦᳓⋧㧩500PL㧦੃ൻߩ᭽ሶ߿ᓧࠄࠇߚᶧṢᓘߥߤߦߟ޿ߡႎ๔ߔࠆ㧚

20PL ߣߒߚ㧚ᴤ⋧ߦ⩿⒳ᴤ㧔࠽ࠞ࡜ࠗ࠹ࠬࠢ㧕ࠍ↪޿㧘ߎߐࠄߦ㧘ߎߎߢ␜ߒߚ㔚⇇ࠍ↪޿ߚᣇᴺ߇㧘↢‛ቇ⊛⹜ᢱߎߦ⇇㕙ᵴᕈ೷ߢ޽ࠆ Tween 80㧔Sigma㧕ࠍ 1㧑㧔v/v㧕ട߃ߦᓇ㗀ࠍਈ߃ࠆ߆ߤ߁߆ࠍ⺞ߴࠆߚ߼ߦ㧘PCR ࠍⴕߞߡ㧘ߘߚ߽ߩߣ㧘ട߃ߥ޿߽ߩࠍ↪ᗧߒߚ㧚᳓ṁᶧߦߪ㧘Milli-Q ᳓ߩᓇ㗀ࠍᬌ⸛ߒߚ㧚㧔⿥⚐᳓㧕ߣ 10mM KCl ṁᶧࠍ↪޿ߚ㧚ࠟ࡜ࠬࠠࡖࡇ࡜࡝࡯㧔ౝᓘ 0.5mm㧘ᄖᓘ 1mm㧘G-1㧘

2.

㕒㔚੃ൻ

Narishige㧕ߩਛߦ࠲ࡦࠣࠬ࠹ࡦ✢㧔0.3mm1㧕ࠍዉ౉ߒߚ߽ߩ㔚⇇ࠍ↪޿ࠆᣇᴺߣߒߡ㧘㔚⇇ࠍශടߔࠆߚ߼ߩ㔚ᭂࠍኈࠍ㜞㔚࿶㔚ᭂߣߒߡ↪޿ߚ㧔࿑ 1-A㧕㧚߹ߚ㧘ࠟ࡜ࠬࠠࡖࡇ࡜ེߩౝㇱ㧘޽ࠆ޿ߪᄖㇱߦ⸳⟎ߒߚ႐วߩ 2 ⒳㘃ߩᣇᴺࠍ⹜࡝࡯ߩవ┵㧔ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩᐩㇱ஥㧕ߪࠪ࡝ࠦ࡯ࡦࠪ࡯ߺߚ㧚ኈེߦߪࡊ࡜ࠬ࠴࠶ࠢ⵾ࡑࠗࠢࡠ࠴ࡘ࡯ࡉࠍ↪޿ߚ㧚࡜ࡦ࠻㧔KE-45T㧘ା⿧ࠪ࡝ࠦ࡯ࡦ㧕ߢኽශߒߚ㧚㔚ᭂߦߎߩ㔚ᭂࠍౝㇱߦ⸳⟎ߔࠆ႐วߪ㧘ኈེౝㇱߦ㜞㔚࿶㔚ᭂߢ޽ࠆࠃ߁ߥᒻ⁁ࠍ↪޿ߚߩߪ㧘㊄ዻࠍࠟ࡜ࠬߢⷒ߁ߎߣߢ㧘㔚࿶㊄ዻ✢ࠍ㈩⟎ߒ㧘ࡑࠗࠢࡠ࠴ࡘ࡯ࡉࠍ⸳⟎ߒߡ޽ࠆࠕ࡞ࡒ࠾ශടਛߩ㔚ᭂ෻ᔕ㧔᳓ߩ㔚᳇ಽ⸃߿㊄ዻ᧚ᢱߩṁ಴㧕ࠍᛥ߃࠙ࡓ⵾࠴ࡘ࡯ࡉ࡜࠶ࠢࠍធ࿾㔚ᭂߣߒߚ㧚ᄖㇱߦ⸳⟎ߔࠆ႐ࠆߚ߼ߢ޽ࠆ㧚วߪ㧘ࡊ࡜ࠬ࠴࠶ࠢ⵾ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩᄖㇱߩ 2 ▎ᚲߦࠕࡑࠗࠢࡠ࠴ࡘ࡯ࡉౝߦ⹜ᢱṁᶧࠍ౉ࠇ㧘࿑ 1-B ߦ␜ߔࠃ߁
ߦ㜞㔚࿶㔚ᭂࠍ⸳⟎ߒ㧘ߘߩࡑࠗࠢࡠ࠴ࡘ࡯ࡉࠍࠕ࡞ࡒ࠾࠙
ࡓࡉࡠ࠶ࠢ㧔CHT100-CB㧘Iwaki㧕ߦ⸳⟎ߒߚ㧚߹ߚ㧘ߎߩࠕ࡞ࡒࡉࡠ࠶ࠢࠍធ࿾㔚ᭂߣߒߚ㧚ߎߩ⁁ᘒߢ㜞㔚࿶㔚ᭂߦࠗࡦࡃ࡯࠲ࡀࠝࡦ࠻࡜ࡦࠬ㧔M-5㧘࡟ࠪ࠶ࡊ㧕ࠍ↪޿ߡ㧘੤ᵹ㜞㔚࿶㧔߅ࠃߘ 17kHz㧕ࠍශടߒߚ㧚㔚࿶୯ߪ 1 ᰴ஥ߦធ⛯ߒߚࠬ࡜ࠗ࠳࠶ࠢ㧔S-130-10㧘ጊ⪉㔚ᯏ㧕ߢ⺞ᢛߒߚ㧚੃ൻᓟߩ᳓Ṣߪ㧘ᱜ┙ဳ㗼ᓸ㏜㧔Axioplan㧘Carl Zeiss㧕ࠍ↪޿ߡⷰኤߒ㧔20 ୚ኻ‛࡟ࡦ࠭㧕㧘᠟ᓇߒߚ↹௝߆ࠄ᳓Ṣᓘࠍ⸘᷹ߒߚ㧚(A)߹ߚ㧘੃ൻߩ᭽ሶࠍⷰኤߔࠆߚ߼ߦ㧘᳓㆏᳓ࠍ౉ࠇߚࡆ࡯ࠞ࡯ߦ㜞㔚࿶㔚ᭂࠍ⸳⟎ߒߚࡑࠗࠢࡠ࠴ࡘ࡯ࡉࠍᶋ߆ߴߡ㧘᳓㆏᳓ࠍធ࿾ߒ㧘㔚࿶ࠍශടߒߡ੃ൻߐߖࠆታ㛎߽ⴕߞߚ㧚ߎߩ੃ൻߩ᭽ሶߪ㧘࠺ࠫ࠲࡞ࡑࠗࠢࡠࠬࠦ࡯ࡊ㧔VH-5000㧘
Keyence㧕ࠍ↪޿ߡⷰኤߒߚ㧚(B)
࿑ 1 ታ㛎ⵝ⟎㧔ౝㇱ㔚ᭂ㧕㧔A㧕᭴ᚑⷐ⚛㧘
㧔B㧕⸳⟎ߩ᭽ሶ
Fig.1 Experimental setup in the case of internalelectrode.
(A) Components, (B) Experimental setup.

࿑ 2 ታ㛎ⵝ⟎㧔ᄖㇱ㔚ᭂ㧕
Fig.2 Experimental setup in the case of external electrodes.

28

30
2.1.2

12006

ᄖㇱ㔚ᭂ

ᄖㇱ㔚ᭂߣߒߡࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩ 2 ▎ᚲߦࠕ࡞ࡒ࠾࠙ࡓ
࠹࡯ࡊࠍᏎ޿ߚ㧚᳓ṁᶧߦߪ↢‛ቇ⊛⹜ᢱ߳ߩᓇ㗀ࠍ⷗ࠆߚ߼ߦ PCR ṁᶧࠍ↪޿ߚ㧚ࡊ࡜ࠬ࠴࠶ࠢ⵾ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߦ㧘0.2mL ࡑࠗࠢࡠ࠴ࡘ࡯ࡉ㧔PCR-02-LC㧘Axygen㧕ࠍ↪޿㧘⇇㕙ᵴᕈ೷ Tween 80㧔1㧑v/v㧕ࠍ฽߻⩿⒳ᴤ 100PL㧘PCR ṁᶧ㧔⚵ᚑߪᓟㅀ㧕5PL ࠍ⹜(A)ᢱṁᶧߣߒߚ㧚ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩᄖ஥ߦ࿑ 2 ߩࠃ߁ߦࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩᐩㇱߣ⹜ᢱṁᶧߩᶧ㕙ߦࠕ࡞ࡒ࠹࡯ࡊࠍᏎ޿ߡ㔚ᭂߣߒߚ㧚ᐩㇱߩ㔚ᭂߦࠗࡦࡃ࡯࠲ࡀࠝࡦ࠻࡜ࡦࠬࠍធ⛯ߒߡ㜞㔚࿶ࠍශടߒߡ㧘਄ㇱߩ㔚ᭂࠍធ࿾ߒߚ㧚߹ߚ㧘੃ൻߩ᭽ሶߪ㧘࠺ࠫ࠲࡞ࡑࠗࠢࡠࠬࠦ࡯ࡊࠍ↪޿ߡⷰኤߒߚ㧚੃ൻᓟߩ᳓Ṣߪ㧘਄ㅀߒߚࠃ߁ߦ㧘ᱜ┙ဳ㗼ᓸ㏜ࠍ↪޿ߡⷰኤߒ㧘᠟ᓇߒߚ↹௝߆ࠄ᳓Ṣᓘࠍ⸘᷹ߒߚ㧚
2.2⚿ᨐߣ⠨ኤ

2.2.1ౝㇱ㔚ᭂ⩿⒳ᴤ㧔500PL㧕ߣ᳓ṁᶧ㧔20PL㧕ࠍࡑࠗࠢࡠ࠴ࡘ࡯ࡉౝߦዉ౉ߒ㧘㕒⟎ߒߚ⁁ᘒߢ੤ᵹ㜞㔚࿶㧔⚂ 17kHz㧕ࠍශടߒߚ㧚㔚࿶ශടߣหᤨߦ᳓ṁᶧ߇ᴤਛߦಽᢔߒᆎ߼㧘ᦨ⚳⊛ߦߪో૕߇⊕Ộߒߚ㧚࿑ 3 ߪ㧘⩿⒳ᴤߣ Milli-Q ᳓ࠍ↪޿ߡ㧘(B)࿑ 4 ੃ൻߩ᭽ሶ 1
㧔A㧕Milli-Q ᳓㧘㧔B㧕KCl ᳓ṁᶧ㧚⇇㕙ᵴᕈ೷㧔1㧑㧕ࠍ฽߻⩿⒳ᴤࠍ૶↪㧚Ꮐ߆ࠄ㔚࿶ශടᤨ㑆߇ 1㧘3㧘5㧘
10 ಽ㧚
Fig.4 Photographs of W/O emulsion 1.
(A) Milli-Q water, (B) KCl solution. Rapeseed oil including
surfactant (1%) was used. voltage applying time (left toright) was 1, 3, 5 and 10 min, respectively.

7kV0-P ߩ੤ᵹ㔚࿶ࠍශടߒߚߣ߈ߩ੃ൻߩ᭽ሶߢ޽ࠆ㧚㔚࿶ࠍශടߔࠆߣ㔚ᭂߩ๟ࠅ߇੃ൻߒᆎ߼㧔࿑ 3-B㧕
㧘10 ⑽ᓟߦ

(A)
(A)

(B)

(B)

(C)

࿑ 3 ੤ᵹ㔚⇇ߦࠃࠆ੃ൻߩ᭽ሶ
㧔A㧕࠮࠶࠻ࠕ࠶ࡊ㧘
㧔B㧕㔚࿶ශട 3 ⑽ᓟ㧘㧔C㧕㔚࿶ශട 10 ⑽ᓟFig.3 Emulsification by AC electric field.
(A) Setup, (B) after 3 seconds and (C) 10 seconds.

࿑ 5 ੃ൻߩ᭽ሶ 2
㧔A㧕Milli-Q ᳓㧘㧔B㧕KCl ᳓ṁᶧ㧚⇇㕙ᵴᕈ೷㧔1㧑㧕ࠍ฽߹ߥ޿⩿⒳ᴤࠍ૶↪㧚Ꮐ߆ࠄ㔚࿶ශടᤨ㑆߇ 1㧘3㧘
5㧘10 ಽ㧚Fig.5 Photographs of W/O emulsion 2.
(A) Milli-Q water, (B) KCl solution. These were in the caseof without surfactant. Voltage applying time (left to right)
was 1, 3, 5 and 10 min, respectively.

29ㅀߴࠆ㧚ශട㔚࿶୯ࠍ 7kV0-P ߦ࿕ቯߒߡ㧘㔚࿶ශടᤨ㑆㧘⇇ߪ߶߷ో૕߇⊕Ộൻߒߚ㧔࿑ 3-C㧕㧚⺃㔚ᕈᶧ૕ߩਛߦዉ㔚ᕈߩ᳓Ṣ㧘޽ࠆ޿ߪዉ㔚ᕈߩዊߐߥ㕙ᵴᕈ೷㧘᳓ṁᶧߩዉ㔚₸ࠍᄌൻߐߖߚߣ߈ߩ⹜ᢱṁᶧߩ᭽᳓Ṣ߇ሽ࿷ߔࠆߣ߈㧘ߘߎߦ㔚⇇ࠍශടߔࠆߣ᳓Ṣߩ⴫㕙߇

ሶࠍ࿑ 4 ߅ࠃ߮࿑ 5 ߦ␜ߔ㧚࿑ 4 ߢߪ㧘⇇㕙ᵴᕈ೷ࠍ฽߻⩿㔚⇇߆ࠄജࠍฃߌߡ㧘ߘߩ᳓Ṣ߇㔚⇇ᣇะߦᒁ߈િ߫ߐࠇߡ⒳ᴤࠍ↪޿㧘࿑ 5 ߢߪ㧘⇇㕙ᵴᕈ೷ࠍ฽߹ߥ޿⩿⒳ᴤࠍ↪޿ᬦ౞ᒻߦߥߞߚࠅ㧘ߘߩᬦ౞ߩవ┵߆ࠄዊߐߥᶧṢߦಽⵚߒߚ㧚᳓ṁᶧߦߪ Milli-Q ᳓㧔࿑ 4-A㧘࿑ 5-A㧕ߣ 10mM KCl

ߚࠅߔࠆߣ޿߁⃻⽎߇㧘ታ㛎⊛ߦ߽ℂ⺰⊛ߦ߽ႎ๔ߐࠇߡ޿

᳓ṁᶧ㧔࿑ 4-B㧘࿑ 5-B㧕ࠍ↪޿㧘㔚࿶ශടᤨ㑆ࠍ 1㧘3㧘5㧘5)

ࠆ 㧚ᶧṢߩಽⵚߪ㧘⇇㕙ᒛജߣ㔚⇇߆ࠄߩജߩࡃ࡜ࡦࠬߦ

10 ಽߣᄌൻߐߖߚ㧚⇇㕙ᵴᕈ೷ߩ᦭ή㧔࿑ 4㧦᦭㧘࿑ 5㧦ή㧕ࠍᲧߴࠆߣ㧘⇇

ࠃߞߡᡰ㈩ߐࠇࠆ㧚ߎߎߢ⷗ࠄࠇߚ੃ൻ⃻⽎ߪ㧘ߎߩ㔚⇇ߦࠃࠆᶧṢߩಽⵚߣ㕙ᵴᕈ೷߇౉ߞߡ޿ࠆ߶߁㧘ߔߥࠊߜ⇇㕙ᒛജ߇ዊߐߥ߶߁ห᭽ߢ޽ࠆߣ⠨߃ࠄࠇࠆ㧚㔚⇇ࠍශടߔࠆߎߣߢ㧘ࡑࠗࠢࡠ߇ࠃࠅဋ৻ߦ੃ൻߒߡ޿ࠆߎߣ߇ࠊ߆ࠆ㧚߹ߚ㧘⇇㕙ᵴᕈ೷࠴ࡘ࡯ࡉᐩㇱߩ᳓ṁᶧߣᴤ⋧ߩႺ⇇㕙ߢ߹ߕᶧṢߩྃ಴߇ࠍ฽߹ߥ޿႐ว㧔࿑ 5㧕ߢ㧘Milli-Q ᳓㧔࿑ 5-A㧕ߣ KCl ᳓ṁ⿠ߎࠅ㧘ߘߩྃ಴ߐࠇߚᶧṢ߇ߐࠄߦ㔚⇇ਛߢᓸዊൻߒߡ޿

ᶧ㧔࿑ 5-B㧕ࠍᲧߴࠆߣ㧘KCl ᳓ṁᶧߩ߶߁߇ࠃࠅဋ৻ߦߥ

ࠆߣ⠨߃ࠄࠇࠆ㧚એ೨ߩႎ๔ߢ㧘੹࿁↪޿ߚࠗࡦࡃ࡯࠲ࡀࠝ

ߞߡ޿ߚ㧚․ߦ㧘ශടᤨ㑆 1 ಽߢߪߘߩ੃ൻ⁁ᘒߩ㆑޿߇㗼ࡦ࠻࡜ࡦࠬߦࠃߞߡ⊒↢ߔࠆ੤ᵹ㔚⇇㧔17kHz㧕ࠍ↪޿ߚ႐⪺ߢ޽ߞߚ㧚৻ᣇ㧘⇇㕙ᵴᕈ೷߇޽ࠆ႐วߪ㧘Milli-Q ᳓㧔࿑วߪ㧘ᶧṢ߇޽߹ࠅᒁ߈િ߫ߐࠇߕߦ㧘ᓸዊߥᶧṢߦಽⵚߒ

4-A㧕ߣ KCl ᳓ṁᶧ㧔࿑ 4-B㧕ߣߩ㑆ߦ㗼⪺ߥ㆑޿߇⷗ࠄࠇ6)ߡ޿ߊߣ޿߁ߎߣࠍⷰኤߒߡ߅ࠅ 㧘ᧄᴺߦ߅޿ߡ߽ห᭽ߩߥ߆ߞߚ㧚⃻⽎߇⿠ߎߞߡ޿ࠆߣ⠨߃ࠄࠇࠆ㧚߹ߚ㧘੃ൻߩೋᦼᲑ㓏ߦߎࠇࠄߩ⚿ᨐ߆ࠄ㧘㔚⇇ࠍ↪޿ߡ੃ൻߒߚ႐ว㧘⇇㕙ᵴᕈ߅޿ߡ㧘㔚ᭂߩ๟ࠅ߇⊕Ộߔࠆߎߣߦ㑐ߒߡߪ㧘㔚ᭂߩᒻ⁁೷ࠍ౉ࠇߥߊߡ߽੃ൻ߇ⴕ߃ࠆߎߣ㧘⇇㕙ᵴᕈ೷߇ή޿႐ว

߆ࠄߘߩㇱಽ߇ࠃࠅᒝ޿㔚⇇ࠍᒻᚑߒߡ޿ࠆߚ߼ߢ޽ࠆߣ

㧔⇇㕙ᒛജ߇ᄢ߈ߥ႐ว㧕ߪ㧘᳓ṁᶧߩዉ㔚₸ߦᓇ㗀ߐࠇࠆ⠨߃ߡ޿ࠆ㧚

ߎߣ߇ࠊ߆ߞߚ㧚᭽‫᧦ߦޘ‬ઙࠍᄌ߃ߚߣ߈ߩ੃ൻߩ᭽ሶߣᶧṢᓘߦߟ޿ߡ

⴫ 1 ߦ࿑ 4㧘࿑ 5 ߩࠛࡑ࡞࡚ࠫࡦࠍ㗼ᓸ㏜ⷰኤߒߡ⸘᷹ߒߚᶧṢᓘࠍ␜ߔ㧚ශടᤨ㑆߇⍴ߊ㧘ో૕߇੃ൻߒߡ޿ߥ޿⹜
ᢱߢߪ㧘⊕Ộߒߡ޿ࠆㇱಽࠍࠨࡦࡊ࡝ࡦࠣߒߚ㧚⴫ 1 ᶧṢᓘ 1

ో ૕ ࠍ ㅢ ߒ ߡᄌ േ ଥ ᢙ ߇ ᄢ߈ ߊ 㧘 CV 㧔 Coefficient ofTable 1 Diameter of the droplets 1.
Time

Diameter*

CV

[min]

[Pm]

[%]

Variance㧕୯߇ 40㨪60%ߦߥߞߚ㧚 ශടᤨ㑆߇ 1 ಽߣ⍴޿႐

Condition
Rapeseed oilWaterวߪ㧘ᶧṢᓘ߇ઁߩ᧦ઙߦᲧߴߡᄢ߈ߊ㧘CV ୯߽ᄢ߈޿㧚3ಽએ਄㔚࿶ࠍශടߒߚ႐วߪ㧘ߘࠇߙࠇߩ᧦ઙߢ㧘ᶧṢᓘ߇

1

8.58r4.33 50.4

3

6.98r3.58 51.3

5

7.33r4.13 56.4

10

8.07r3.86 47.9

1

7.43r4.52 60.8

3

7.32r4.09 55.9

5

7.94r3.65 46.0

10

6.25r2.87 45.9

1

7.31r4.14 56.6

3

6.69r3.14 46.9

5

7.08r2.93 41.3

Applied voltage

Diameter

106.43r2.78 43.2

[kV0-P][Pm]

18.82r4.01 45.4

4

9.44r4.3746.3

36.71r2.60 38.7

69.83r4.25

43.3

57.44r3.00 40.4

88.74r4.54

51.9

107.38r3.27 44.4

1010.84r4.08

37.6*MeanrSD.

ᄢ߈ߊᄌࠊࠆߎߣ߇ߥ߆ߞߚ㧚
1% Tween 80

Milli-Q⴫ 2 ߦශട㔚࿶ߩᄢ߈ߐࠍᄌൻߐߖߚߣ߈ߩᶧṢᓘࠍ␜ߔ㧚ߎߩߣ߈㧘㔚࿶ශടᤨ㑆ߪ 5 ಽߣߒ㧘1%Tween 80 ࠍ฽߻⩿⒳ᴤ㧔500PL㧕ߣ Milli-Q ᳓㧔20PL㧕ࠍ↪޿ߚ㧚ශട㔚࿶ࠍ 4㧘6㧘8㧘10kV0-P ߣᄌൻߐߖߚ߇ᶧṢᓘߦᄢ߈

1% Tween 80

10mM KClߥᄌൻߪߥ߆ߞߚ㧚ߒ߆ߒ㧘10kV0-P ࠍශടߒߚ႐วߪઁߩ႐
วߦᲧߴߡ CV ୯߇ૐ߆ߞߚ㧚߹ߚ㧘4kV0-P ߩߣ߈ߪ㧘࿑ 5-A
⴫ 2 ᶧṢᓘ 2

No surfactant

No surfactant

Milli-Q

10mM KCl

Table 2 Diameter of the droplets 2*MeanrSD.

CV [%]

3030ߩශടᤨ㑆 1 ಽߣหߓࠃ߁ߦ߹߫ࠄߦ੃ൻߒߡ޿ߚ㧚12006

ߐ߇ 0.3㨪0.5mm ߢ޽ߞߚ㧚ᄖ஥ߦ㔚ᭂࠍ㈩⟎ߒߚߣ߈ߩ੃

⴫ 1 ߣ⴫ 2 ߩ⚿ᨐࠃࠅ㧘ߎߎߢ↪޿ߚ᧦ઙߩ㔚࿶ශടᤨ㑆ൻߩ᭽ሶࠍ࿑ 6 ߦ␜ߔ㧚㔚࿶ࠍශടߔࠆߣ㧘ࡑࠗࠢࡠ࠴ࡘ࡯߅ࠃ߮㔚࿶ߩᄢ߈ߐߣᶧṢᓘߦ㗼⪺ߥ㑐ଥ߇ߥ޿ߣ޿߁ߎࡉᐩㇱߩ᳓ṁᶧ߇ធ࿾㔚ᭂ஥ߦಽᢔߒߡ޿ߊ᭽ሶ߇⏕⹺ߐߣ߇ࠊ߆ߞߚ㧚ߟ߹ࠅ㧘4㨪10kV0-P ߩ▸࿐ߢߪᶧṢᓘ߇ᄢ߈ࠇߚ㧚ߎߩߣ߈㧘ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩౝოㇱߩ᳓ṁᶧߣᴤ⋧ߊᄌࠊࠄߥ޿ߣ޿߁ߎߣ߇⸒߃ࠆ㧚ߎߎߢᓧࠄࠇߚᶧṢࠃࠅߣߩ⇇㕙߆ࠄዊߐߥ᳓Ṣ߇ធ࿾㔚ᭂ㧔਄ㇱ㧕ߦะ߆ߞߡಽᢔ߽ᄢ߈ߥᶧṢࠍᓧࠆߚ߼ߦߪ㧘ࠃࠅዊߐߥ㔚࿶ࠍ㧘߹ߚ㧘ዊߒߡ޿ߞߚ㧚ዉ૕ߢ޽ࠆ᳓ṁᶧ㧔Milli-Q ᳓㧘ዉ㔚₸㧦ߐߥᶧṢࠍᓧࠆߚ߼ߦߪ㧘චಽߦᄢ߈ߥ㔚࿶ࠍශടߔࠆᔅⷐ

1.47PS/cm㧕ౝߪห㔚૏ߢ޽ࠆߣ⠨߃ࠄࠇࠆߚ߼㧘᳓㧙ᴤ⇇㕙߇޽ࠆߣ⠨߃ࠄࠇࠆ㧚ߚߛߒ㧘࿑ 4 ߣ࿑ 5 ߩ࿑ߩᲧセ߆ࠄ㧘ߦ․ቯߩ㔚૏߇⺃ዉߐࠇࠆߣ⠨߃ࠄࠇࠆ㧚ߎߩߣ߈㧘᳓㧙ᴤ⇇㕙ᒛജߩ㆑޿ߦࠃߞߡో૕ߩ⊕Ộᐲ߇ᄌࠊߞߡ޿ࠆߎߣ

⇇㕙ߪࡑࠗࠢࡠ࠴ࡘ࡯ࡉౝߩ౞㍙⁁ߩㇱಽߢᒻᚑߐࠇߡ޿߆ࠄ㧘ࡑࠗࠢࡠ࠴ࡘ࡯ࡉᐩㇱߦᴉࠎߢ޿ࠆ᳓ṁᶧㇱ㧔20PL㧕ࠆߩߢ㧘᳓́ᴤ⇇㕙ߩᄖ✼ㇱ㧔ࡑࠗࠢࡠ࠴ࡘ࡯ࡉߦធߒߡ޿߆ࠄᶧṢࠍಽᢔߐߖࠆߚ߼ߦߪ㧘㔚࿶୯ߩᄢ߈ߐ߿ශടᤨ㑆ࠆㇱಽ㧕ߩ㔚⇇߇ࠛ࠶ࠫߩലᨐߦࠃߞߡਛᔃㇱಽߦᲧߴߡᄢ߇ᓇ㗀ߒߡ޿ࠆߣ⠨߃ࠄࠇࠆ㧚߈ߊߥࠆ㧚ߎߩ㔚⇇ᒻᚑߪ㔚⇇⸘▚࠰ࡈ࠻㧔Maxwell SV㧕ࠍߎߎߢᓧࠄࠇߚᶧṢߪ㧘ᄌേଥᢙ߇㕖Ᏹߦᄢ߈ߊනಽᢔߣ↪޿ߚ㔚⇇⸘▚ߩ⚿ᨐ߆ࠄ߽⏕⹺ߐࠇߡ޿ࠆ㧚ߎߩ㔚⇇ߦࠃߪ⸒߃ߥ޿㧚਄ㅀߒߚࠃ߁ߥ W/O ࠛࡑ࡞࡚ࠫࡦࠍ↢ൻቇ෻ߞߡ㧘ౝㇱߦ㔚ᭂࠍ⟎޿ߚ႐วߣหߓࠃ߁ߦ㧘᳓ṁᶧㇱಽ߆ᔕ♽ߦ↪޿ࠆ႐วߪ㧘෻ᔕߩဋ৻ᕈࠍ଻⸽ߔࠆߚ߼ߦ㧘ᒻᚑࠄᶧṢ߇ྃ಴ߒߡ޿ࠆߣ⠨߃ࠄࠇࠆ㧚ߎߎߢᓧࠄࠇߚᶧṢᓘߐࠇࠆᶧṢߪනಽᢔߢ޽ࠆߎߣ߇ᦸ߹ߒ޿㧚ߘߩߚ߼㧘੹ᓟߩಽᏓߪ㧘ᓟㅀߩ⴫ 3 ߦ␜ߒߡ޽ࠆ㧚ߩᡷ⦟߇ߐࠄߦᔅⷐߢ޽ࠆ㧚ᬌ⸛⺖㗴ߣߒߡߪ㧘㔚ᭂᒻ⁁߿ߩߢ㧘ᡬᜈ૞ᬺߦࠃࠆᳪᨴ‛⾰ߩᷙ౉ࠍోߊ⠨߃ߥߊߡ߽⦟๟ᵄᢙଐሽᕈߥߤ߇᜼ߍࠄࠇࠆ㧚
2.2.2ᄖㇱ㔚ᭂߦࠃࠆ੃ൻߪ㧘⹜ᢱṁᶧߣᡬᜈㇱಽ߇ធ⸅ߒߥ޿ᄖㇱ㔚ᭂ޿ߣ޿߁․ᓽ߇޽ࠆ㧚⴫ 3 ߦ޽ࠆࠃ߁ߦ㧘ౝㇱߦ㔚ᭂࠍ⟎޿

࿑ 2 ߦ␜ߔࠃ߁ߦࡑࠗࠢࡠ࠴ࡘ࡯ࡉ࠴ࡘ࡯ࡉߩᄖ஥ߦ㔚ᭂࠍ㈩⟎ߒߡ߽ࡑࠗࠢࡠ࠴ࡘ࡯ࡉౝߢ੃ൻߔࠆ᭽ሶ߇⏕⹺ߐߚ႐วߣห᭽ߦ㧘ᄌേଥᢙ߇ᄢ߈޿ߎߣ߇੹ᓟߩᬌ⸛⺖㗴ߢ޽ࠆ㧚ࠇߚ㧚ߎߎߢ↪޿ߚ 0.2mL ࠴ࡘ࡯ࡉߪࡐ࡝ࡊࡠࡇ࡟ࡦ⵾ߢෘ
3.

PCR ߳ߩᔕ↪

ߎࠇ߹ߢߦ㧘W/O ࠛࡑ࡞࡚ࠫࡦࠍ೑↪ߒߚ 1 ಽሶߩ㍌ဳ
DNA ߆ࠄߩ PCR Ⴧ᏷ߦᚑഞߒߡ޿ࠆ 2)㧚ߎࠇߪ㧘㍌ဳ DNAࠍᓸዊߥᶧṢߩਛߦዉ౉ߔࠆߎߣߢ㧘㍌ဳ DNA Ớᐲࠍ╬ଔ⊛ߦ㜞Ớᐲߦߒ㧘㜞ല₸ߦ෻ᔕࠍⴕߞߚߚ߼ߢ޽ࠆ㧚ߎߎߢߪ㧘ᧄ੃ൻᣇᴺࠍ W/O ࠛࡑ࡞࡚ࠫࡦࠍ↪޿ߚ PCR ᴺ߳ㆡ↪
ߒ㧘ߘߩᓇ㗀ࠍ⺞ߴߚ㧚
3.1ታ㛎ᣇᴺPCR ṁᶧߩ⚵ᚑߪ㧘PCR ࡃ࠶ࡈࠔ࡯㧔20mM Tris-HCl 㧔pH(A)8.8㧕
㧘2mM MgSO4㧘10mM KCl㧘10mM (NH4) 2SO4㧘0.1% Triton
X-100㧘0.1mg/mL BSA㧕
㧘0.2mM dNTPs㧔࠲ࠞ࡜ࡃࠗࠝ㧕
㧘10mMDTT 㧔 dithiothreitol 㧘 ๺ శ ⚐ ⮎ 㧕㧘 0.2PM Primer1
㧔5’-CTTGAGTCCAACCCGGTAAG 㧕㧔ࡈ࠽ࠦࠪ㧕㧘0.2PM
Primer2㧔5’-GGGGAGTCAGGCAACTATGG㧕㧔ࡈ࠽ࠦࠪ㧕㧘
pUC19 DNA㧔5ng㧕㧔࠲ࠞ࡜ࡃࠗࠝ㧕㧘2.5U PfuTurbo DNA
Polymerase 㧔Stratagene㧕ߣߒߚ‫ߩߎޕ‬෻ᔕߦࠃߞߡ㧘522bp
ߩჇ᏷↥‛ࠍᓧࠆߎߣ߇ߢ߈ࠆ‫ޕ‬ታ㛎ߦߪ 0.2mL ࠴ࡘ࡯ࡉࠍ(B)(C)࿑ 6 ੃ൻߩ᭽ሶ㧔ᄖㇱ㔚ᭂ㧕㧔A㧕࠮࠶࠻ࠕ࠶ࡊ㧘
㧔B㧕㔚࿶ශട 3 ⑽ᓟ㧘㧔C㧕10 ⑽ᓟ㧚Fig.6 Emulsification using external electrode.
(A) Experimental setup, (B) after 3 seconds and (C) 10
seconds.

↪޿ߡ㧘ౝㇱ㔚ᭂߣᄖㇱ㔚ᭂߩ 2 ⒳㘃ߢ੃ൻࠍⴕߞߚ㧚੃ൻࠍⴕߞߚᓟ㧘95͠㧘3 ಽߩᓟߦ 95͠㧘30 ⑽㧧60͠㧘30 ⑽㧧
72͠㧘1 ಽࠍ 12 ࠨࠗࠢ࡞ⴕ޿㧘16000g ߢ 1 ಽ㑆㧔4͠㧕㆙ᔃᠲ૞ࠍⴕ޿㧘ࠛࡑ࡞࡚ࠫࡦࠍ⎕უߒ㧘2 ጀߦಽ㔌ߒߚᓟ㧘

31

ߐࠄߦ PCR ࠍ 20 ࠨࠗࠢ࡞ⴕߞߚ‫ޕ‬PCR ↥‛ߪ 0.8%ࠕࠟࡠ࡯ࠬࠥ࡞ߢ㔚᳇ᵒേߒߡ⏕⹺ߒߚ‫ޕ‬
3.2⚿ᨐߣ⠨ኤ

⴫ 3 ߦ㧘
ߎߎߢᬌ⸛ߒߚฦ᧦ઙߦ߅ߌࠆᶧṢᓘࠍ⸥タߔࠆ㧚ߎߎߢ㧘޿ߕࠇߩ᧦ઙߦ߅޿ߡ߽ 1 ಽએౝߦචಽߦో૕߇⊕Ộߒߡ޿ߚߩߢ㧘㔚࿶ශടᤨ㑆ࠍᦨᄢߢ 1 ಽ㑆ߣߒߚ㧚߹ߚ㧘㐳ᤨ㑆ߩ㔚࿶ශടߦࠃࠆ㉂⚛߳ߩᓇ㗀ࠍᛥ೙ߔࠆߚ߼ߢ߽
޽ࠆ㧚5kV0-P㧘15 ⑽㧔࿑ 7-lane2㧕㧧3kV0-P㧘15 ⑽߅ࠃ߮ 30 ⑽㧔࿑ 7-lane3㧘4㧕㧘ߩߣ߈ߪో૕߇ቢోߦ⊕Ộߒߡ޿ߥ߆ߞߚ㧚ౝㇱ㔚ᭂߩ႐ว㧘਄ㅀߩ 1.5mL ࠴ࡘ࡯ࡉࠍ↪޿ߚ႐วߣᲧߴ
ࠆߣ㧘ᶧṢᓘߪ߅ࠃߘඨಽߣߥߞߚ㧚ߎࠇߪ㧘ࡑࠗࠢࡠ࠴ࡘ

࿑ 7 PCR ߩ⚿ᨐ㧔㔚᳇ᵒേ౮⌀㧕࡟࡯ࡦ M㧦ಽሶ㊂ࡑ࡯ࠞ࡯㧔O/Hind III㧕㧘࡟࡯ࡦ 1㧦ࡐࠫ࠹ࠖࡉࠦࡦ࠻ࡠ࡯࡞㧔ࠛࡑ࡞࡚ࠫࡦߥߒ㧕㧘࡟࡯ࡦ
2-8㧦⴫ 3 ߦ⸥ㅀߒߡ޿ࠆ᧦ઙߢ੃ൻߒߚ߽ߩ
Fig.7 The results of the PCR (gel electrophoresis).
Lane M: molecular weight marker (O/Hind III), lane 1:
positive control (without emulsion). Lane 2-8 correspond to
the sample mentioned in Table3.

࡯ࡉࠍ 0.2mL ↪ߦᄌ߃ߚߚ߼㧘ౝㇱߩ㔚⇇߇ᄢ߈ߊߥߞߚ߆
ࠄߢ޽ࠆߣ⠨߃ࠄࠇࠆ㧚ߒ߆ߒ㧘CV ୯߇ 35㨪50㧑ߣ㕖Ᏹߦᄢ߈޿ߚ߼㧘੹ᓟߐࠄߦᡷ⦟߇ᔅⷐߢ޽ࠆ㧚ታ㛎⚿ᨐࠍ࿑ 7 ߦ␜ߔ㧚੃ൻߐߖߡ޿ߥ޿⹜ᢱ㧔࡟࡯ࡦ 1㧕ߣᲧセߔࠆߣ㧘޿ߕࠇߩ᧦ઙߦ߅޿ߡ߽᣿ࠄ߆ߥ㒖ኂߪ⏕⹺ߐࠇߥ߆ߞߚ㧚ߎࠇߦࠃࠅ㧘㔚⇇ࠍ↪޿ߚ੃ൻᴺ߇ PCR ߦ↪
޿ࠆߎߣ߇ߢ߈ࠆߎߣ߇␜ߐࠇߚ㧚ઁߩ↢ൻቇ෻ᔕ߳ߩㆡ↪

4.

߽น⢻ߢ޽ࠆߣ੍ᗐߐࠇࠆ㧚ᧄႎ๔ߦ߅޿ߡ㧘એਅߩ⍮⷗ࠍᓧߚ‫ޕ‬߹ߣ߼ߒ߆ߒ㧘ౝㇱ㔚ᭂߩ႐วߦ߅޿ߡ㧘ශട㔚࿶ࠍ 7kV0-P ߦ਄1㧕 ࡑࠗࠢࡠ࠴ࡘ࡯ࡉౝߦ੤ᵹ㜞㔚࿶㔚ᭂࠍ㈩⟎ߔࠆߎߣ᣹ߐߖࠆߣ↥‛ࠍᓧࠆߎߣ߇ߢ߈ߥ߆ߞߚ㧚ߎߩߣ߈㧘෻ᔕߢ㧘㕒⟎ߒߚ⁁ᘒߩ᳓㧙ᴤᷙวᶧࠍ੃ൻߢ߈ߚ㧚⚳ੌᤨߩ᳓ṁᶧߩ pH ߇㉄ᕈ㧔pH ᜰ␜⮎ߩ⦡߆ࠄ߅ࠃߘ pH32㧕 㜞㔚࿶㔚ᭂࠍࡑࠗࠢࡠ࠴ࡘ࡯ࡉߩᄖ஥ߦ㈩⟎ߒߡ߽੃એਅ㧕ߦߥߞߡ޿ࠆߎߣ߇⏕⹺ߐࠇߚ㧚߹ߚ㧘ߎߩ⚿ᨐߪ PCR
ṁᶧਛߦ 10mM DTT㧔㉄ൻ㒐ᱛ೷㧕ࠍ฽ࠎߢ޿ࠆ߇㧘ߎࠇࠍൻߢ߈ߚ㧚3㧕 㔚⇇ࠍ↪޿ߡ੃ൻߒߡ߽ PCR ߳ߩ㗼⪺ߥᓇ㗀ߪ⷗ࠄࠇᷝടߖߕߦ㧘5kV0-P ߩ㔚࿶ࠍශടߒߚߣ߈ߦ߽Ⴧ᏷↥‛ࠍᓧߥ߆ߞߚ㧚ࠆߎߣ߇ߢ߈ߥ߆ߞߚ㧔ౝㇱ㔚ᭂ࡮ᄖㇱ㔚ᭂߩਔᣇߣ߽㧕㧚੹ᓟߩ⺖㗴ߣߒߡߪ㧘↢ൻቇ෻ᔕ߿ࠦࡦࡆ࠽࠻࡝ࠕ࡞ߥ෻ߎࠇࠄߩߎߣ߆ࠄ㧘↢ൻቇ෻ᔕߦᔕ↪ߔࠆ㓙ߪ㧘ṁᶧߩ pHᔕ߳ᔕ↪ߔࠆ㓙ߪ㧘නಽᢔߥᶧṢ߇ᒻᚑߐࠇࠆߎߣ߇ᔅⷐߢ

ࠍᄌൻߐߖߥ޿ࠃ߁ߥㆡᒰߥ㔚࿶㧔5kV0-P એਅ㧕ࠍㆬ߮㧘㔚޽ࠅ㧘CV ୯ࠍᛥ߃ߡ㧘ᶧṢᓘࠍࠦࡦ࠻ࡠ࡯࡞ߢ߈ࠆࠃ߁ߦ㧘⇇ߦࠃࠆ㉄ൻࠍᛥ೙ߔࠆኻ╷߇ᔅⷐߢ޽ࠆߎߣ߇ࠊ߆ߞߚ㧚๟ᵄᢙᔕ╵ߥߤࠍ᷹ቯߔࠆߎߣࠍ฽߼ᡷ⦟ࠍട߃ࠆᔅⷐ߇޽ࠆ㧚

⴫ 3 ᶧṢᓘಽᏓ㧔PCR ߳ߩᔕ↪㧕
Table 3 Diameter of the droplets of PCR application.
Lane
VoltageTime

Diameter*CV

Position ofin[kV0-P][sec][Pm]%]

electrodeFig.
7515

4.24r2.21

52.2

2

530

3.24r1.15

35.4

3315

3.61r1.84

51.1

330

3.69r1.70

46.0

5360

4.14r2.08

50.2

6560

4.13r2.19

52.9

5.5

30

4.04r1.73

42.8*MeanrSD.

InsideOutside478

ᧄ⎇ⓥߪ․೎⎇ⓥຬᅑബ⾌㧔No.08351㧕߅ࠃ߮ᢥㇱ⑼ቇ⋭⑼ቇ⎇ⓥ⵬ഥ㊄㧔ၮ⋚⎇ⓥ B㧔2㧕15360440㧕ߩഥᚑࠍฃߌߡⴕࠊࠇߚ߽ߩߢ޽ࠅ㧘ߎߎߦ⸥ߒ㧘ᷓߊᗵ⻢޿ߚߒ߹ߔ‫ޕ‬ෳ⠨ᢥ₂
A. Sepp, D.S. Tawfik And A.D. Griffiths: FEBS Lett., 532(2002) 455
2) M. Nakano, J. Komatsu, S. Matsuura, K. Takashima, S.
Katsura and A. Mizuno: J. Biotechnol., 102 (2003) 1173) ੗㊁৻㧘੹⍹ትਯ㧘ቲỈశ♿㧘⮮✽ൎᒾ㧦ൻቇᎿቇ⺰ᢥ㓸㧘18㧔1992㧕8194) ᧲ㄞஜ㧘ᷰㄝ᣽㧦ᣣᧄൻቇળ⹹㧘10㧔1979㧕12875) J.W. Ha and S.M. Yang: J. Fluid Mech., 405 (2000) 1316) ਛ㊁㆏ᒾ㧘㜞ፉ๺ೣ㧘᩵ㅴม㧘᳓㊁ᓆ㧦╙ 28 ࿁㕒㔚᳇ቇળ⻠Ṷ⺰ᢥ㓸’04㧘p.121㧘㕒㔚᳇ቇળ(2004)
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