The aim of this study is to de
v
elop a nucleic acid sensor based on piezoelectric crystal microbalance system (QCM) for following
hybridization. Piezoelectric quartz crystal surfaces were first treated in a glow
/
discharge apparatus with ethylene diamine (EDA)
plasma at 15 W (discharge power), 2.5 min (incubation time) and 35 ml/min (monomer flow rate) to create amino groups on the
crystal surfaces. The thickness of the EDA
/
plasma film formed was about 43
9
/
24 A
̊
. Then, the amino groups on the crystal surfaces
were con
v
erted to aldehyde groups by reacting the amino groups with glutaraldehyde (GA) at different conditions. A GA
concentration of 2.5% and an incubation time of 2 h were selected as optimal
v
alues at this step, corresponding to a GA surface
concentration of about 270 ng/cm
2
. A double strand Oligonucleotides, ha
v
ing one extra base on 5
?
-end of one of the complementary
strands, were immobilized through the amino groups of this base onto the GA-modified crystals. Optimal immobilization conditions
were as follows: oligonucleotide concentration: 1
m
g/ml; time: 3 h; pH: 9.2 carbonate buffer; ionic strength: 0.1; and temperature:
20
8
C. The QCM sensor carrying the co
v
alently bound strand was used in the hybridization experiments, which showed that
equilibrium is achie
v
ed in about 5 min, and the frequency shift measured is related to the concentration of the target strand to be
measured within the medium.
จุดมุ่งหมายของการศึกษาครั้งนี้คือการเดวีElop ดังต่อไปนี้การผสมพันธุ์ พื้นผิวผลึกคริสตัล / อุปกรณ์ปล่อยกับ พลาสม่า พื้นผิวผลึก ความหนาของ / ฟิล์มพลาสม่าที่เกิดขึ้นเป็นประมาณ 9/24 ̊ จากนั้นกลุ่มอะมิโนบนพื้นผิวผลึกเป็นนักโทษวีerted GA ความเข้มข้น ชั่วโมงได้รับเลือกเป็นที่ดีที่สุดโวalues ความเข้มข้นของประมาณ 2 oligonucleotides v ไอเอ็นจีเป็นหนึ่งในฐานพิเศษในวันที่ 5? -End ของหนึ่งในการเสริมเส้นถูกตรึงผ่านกลุ่มอะมิโนที่ฐานนี้ไปยังผลึกGA The aim of this study is to de
v
elop a nucleic acid sensor based on piezoelectric crystal microbalance system (QCM) for following
hybridization. Piezoelectric quartz crystal surfaces were first treated in a glow
/
discharge apparatus with ethylene diamine (EDA)
plasma at 15 W (discharge power), 2.5 min (incubation time) and 35 ml/min (monomer flow rate) to create amino groups on the
crystal surfaces. The thickness of the EDA
/
plasma film formed was about 43
9
/
24 A
̊
. Then, the amino groups on the crystal surfaces
were con
v
erted to aldehyde groups by reacting the amino groups with glutaraldehyde (GA) at different conditions. A GA
concentration of 2.5% and an incubation time of 2 h were selected as optimal
v
alues at this step, corresponding to a GA surface
concentration of about 270 ng/cm
2
. A double strand Oligonucleotides, ha
v
ing one extra base on 5
?
-end of one of the complementary
strands, were immobilized through the amino groups of this base onto the GA-modified crystals. Optimal immobilization conditions
were as follows: oligonucleotide concentration: 1
m
g/ml; time: 3 h; pH: 9.2 carbonate buffer; ionic strength: 0.1; and temperature:
20
8
C. The QCM sensor carrying the co
v
alently bound strand was used in the hybridization experiments, which showed that
equilibrium is achie
v
ed in about 5 min, and the frequency shift measured is related to the concentration of the target strand to be
measured within the medium.
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