XIE/2:2025/Dec: Difference between revisions

Revision as of 08:58, 13 December 2025

12.12

Triplex DNA pH Test

Sample preparation

ssDNA stocks: s1 (3'Cy5), S2, TFO (3'Cy3), S‑S1 (3'Cy5), S‑S2, and S‑TFO (3'Cy3) were each prepared at 1 mM in nuclease-free water.
Buffers: PBS adjusted to pH 5.5, 6.5, 7.0, 7.5, and 8.5; prepare 50 mL of each pH condition.

Incubation

For each pH condition, set up two groups (Group 1 and Group 2). To each reaction, first add 194 µL of the corresponding PBS buffer.

Group 1: add S1, S2, and TFO , 2 µL each.
Group 2: add S-S1, S-S2, and S-TFO , 2 µL each.

Name	Buffer	SsDNA
5.5-1	194 uL, pH 5.5	S1, S2, TFO, 2 uL each
5.5-2	149 uL, pH 5.5	S-S1, S-S2, S-TFO, 2 uL each
6.5-1	194 uL, pH 6.5	S1, S2, TFO, 2 uL each
6.5-2	194 uL, pH 6.5	S-S1, S-S2, S-TFO, 2 uL each
7.0-1	194 uL, pH 7.0	S1, S2, TFO, 2 uL each
7.0-2	194 uL, pH 7.0	S-S1, S-S2, S-TFO, 2 uL each
7.5-1	194 uL, pH 7.5	S1, S2, TFO, 2 uL each
7.5-2	194 uL, pH 7.5	S-S1, S-S2, S-TFO, 2 uL each
8.5-1	194 uL, pH 8.5	S1, S2, TFO, 2 uL each
8.5-2	194 uL, pH 8.5	S-S1, S-S2, S-TFO, 2 uL each

PCR: 37℃ 3h

Fluorescence spectrophotometer

Sample preparation: dilute each sample to 1%.
Measurement channels

IDA channel: excitation 550 nm; emission 670 nm.
IDD channel: excitation 550 nm; emission 570 nm.
IAA channel: excitation 650 nm; emission 670 nm.

@@ Line 1: / Line 1: @@
 <div style="color: black !important; font-family: 'Times New Roman', SimSun, serif !important, line-height:1.5;">
 == 12.12 ==
-*<big>'''<span style="color:red;">Triplex DNA pH Test</span>'''<big>
+*<big>'''<span style="color:red;">Triplex DNA pH Test</span>'''</big>
 === Sample preparation ===
-*ssDNA:s1(3'Cy5)、S2、TFO(3'Cy3)、S-S1(3'Cy5)、S-S2、S-TFO（3'Cy3）各用纯水配成 1mM.
+*ssDNA stocks: s1 (3'Cy5), S2, TFO (3'Cy3), S‑S1 (3'Cy5), S‑S2, and S‑TFO (3'Cy3) were each prepared at 1 mM in nuclease-free water.
-*Buffer:PBS:5.5、6.5、7.0、7.5、8.5(PH) 各 50 ml.
+*Buffers: PBS adjusted to pH 5.5, 6.5, 7.0, 7.5, and 8.5; prepare 50 mL of each pH condition.
 === Incubation ===
-*分组：每个 pH 下设立两组（1、2），每组先各加入194 ul 对应 Buffer。组1，加入 S1、S2、TFO 各 2ul；组 2，加入 S-S1、S-S2、S-TFO 各 2ul。
+*For each pH condition, set up two groups (Group 1 and Group 2). To each reaction, first add 194 µL of the corresponding PBS buffer.
+#Group 1: add S1, S2, and TFO , 2 µL each.
+#Group 2: add S-S1, S-S2, and S-TFO , 2 µL each.
 {| class="wikitable"style="width:80%;"
-! 编号
+! Name
 ! Buffer
 ! SsDNA
@@ Line 14: / Line 17: @@
 | 5.5-1
 | 194 uL, pH 5.5
-| S1, S2, TFO 各 2 uL
+| S1, S2, TFO, 2 uL each
 |-
 | 5.5-2
 | 149 uL, pH 5.5
-| S‑S1, S‑S2, S‑TFO 各 2 uL
+| S-S1, S-S2, S-TFO, 2 uL each
 |-
 | 6.5-1
 | 194 uL, pH 6.5
-| S1, S2, TFO 各 2 uL
+| S1, S2, TFO, 2 uL each
 |-
 | 6.5-2
 | 194 uL, pH 6.5
-| S‑S1, S‑S2, S‑TFO 各 2 uL
+| S-S1, S-S2, S-TFO, 2 uL each
 |-
 | 7.0-1
 | 194 uL, pH 7.0
-| S1, S2, TFO 各 2 uL
+| S1, S2, TFO, 2 uL each
 |-
 | 7.0-2
 | 194 uL, pH 7.0
-| S‑S1, S‑S2, S‑TFO 各 2 uL
+| S-S1, S-S2, S-TFO, 2 uL each
 |-
 | 7.5-1
 | 194 uL, pH 7.5
-| S1, S2, TFO 各 2 uL
+| S1, S2, TFO, 2 uL each
 |-
 | 7.5-2
 | 194 uL, pH 7.5
-| S‑S1, S‑S2, S‑TFO 各 2 uL
+| S-S1, S-S2, S-TFO, 2 uL each
 |-
 | 8.5-1
 | 194 uL, pH 8.5
-| S1, S2, TFO 各 2 uL
+| S1, S2, TFO, 2 uL each
 |-
 | 8.5-2
 | 194 uL, pH 8.5
-| S‑S1, S‑S2, S‑TFO 各 2 uL
+| S-S1, S-S2, S-TFO, 2 uL each
 |}
-*PCR: 10组样品编号，37℃ 3h
+*PCR: 37℃ 3h
-=== 荧光分光光度计 ===
-*样品处理：每个样稀释至1%
+=== Fluorescence spectrophotometer ===
-*测量通道
+*Sample preparation: dilute each sample to 1%.
-#IDA-measure:激发波长 550 nm;发射波长 670 nm
+*Measurement channels
-#IDD-measure:激发波长 550 nm;发射波长 570 nm
+#IDA channel: excitation 550 nm; emission 670 nm.
-#IAA-measure:激发波长 650 nm;发射波长 670 nm
+#IDD channel: excitation 550 nm; emission 570 nm.
+#IAA channel: excitation 650 nm; emission 670 nm.