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Data
Recovery of Trace Amounts of Nucleic Acid
λ/HindIII fragments (10 ng in 500 µL) were precipitated using 1 mL of ethanol in the presence of 0.1 mol/L sodium acetate and analyzed by agarose gel electrophoresis.
Lane 1 : λ/HindIII (Contorol; no precipitation treatment)
Lane 2 : λ/HindIII+Ethachinmate 3 µl (immediate centrifugation)
Lane 3 : λ/HindIII Incubation at -20 °C overnight
Lane 4 : λ/HindIII Incubation at -80 °C for 20 min.
Result
Under the condition of 0.1 mol/L sodium acetate, adding Ethachinmate allows for quantitative recovery of trace amounts of DNA without the need for low-temperature incubation.
Low-Molecular-Weight RNA Ethanol Precipitation
Synthetic 21mer RNA samples (10 ng, 20 ng, 40 ng, and 80 ng) were subjected to ethanol precipitation under the following conditions:
| Sample | Synthesized 21 mer RNA (10 ng,20 ng,40 ng,80 ng) |
|---|---|
| Ethanol precipitation | Each 100 µL RNA solution was added 3M Sodium acetate 3.3 µL and Ethachinmate 1 µL. All samples were immediately precipitated after adding Ethanol. |
| Electrophoresis | 15 % polyacylamide gel |
| RNA Volume | ||
| Lane 1 | Before Ethanol Precipitaion |
10ng |
| Lane 2 | 20ng | |
| Lane 3 | 40ng | |
| Lane 4 | 80ng | |
| Lane 5 | Without Ethachinmate |
10ng |
| Lane 6 | 20ng | |
| Lane 7 | 40ng | |
| Lane 8 | 80ng | |
| Lane 9 | With Ethachinmate |
10ng |
| Lane 10 | 20ng | |
| Lane 11 | 40ng | |
| Lane 12 | 80ng | |
Affect for enzymes
Data 1. Restriction Enzyme Digestion and T4 DNA Ligase
λ DNA was subjected to digestion, ligation, and re-digestion using restriction enzymes (Hae III, Hind III) and T4 DNA Ligase, in the presence or absence of Ethachinmate (1 µL per 10 µL reaction mixture).
| Restriction Enzyme | Hae III | Hind III |
Lane 1: Cut |
||
| Etachinmate | - | + | - | + | |
| Result |  |
 |
 |
 |
|
Result
Ethachinmate did not affect the anzyme activity of restriction ligation efficiency.
Data 2. Reverse Transcriptase
In the presence of Ethachinmate, [3H]dTMP was incorporated into the poly(rA)·(dT)12-8 template using AMV Reverse Transcriptase (RTase).
Reverse Transcription Reaction
50 µL [50 mmol/l Tris-HCl(pH8.3), 6 mmol/l MgCl2, 40 mmol/l KCl, 0.5 mmol/l [3H]dTTP, 0.4 mmol/l poly(rA)・(dT)12-8, 5 units AMV RTase]
Temperature: 42℃
| RTase | Ethachinmate | |
| - | 10 µL | ○-○ |
| + | 0 µL | ●-● |
| + | 5 µL | □-□ |
| + | 10 µL | ■-■ |
Result
Ethachinmate did not affect the activity of Reverse Transcriptase.
Data 3. Taq DNA Polymerase
In the presence of Ethachinmate, a DNA (about 600 bp) was amplified using Taq DNA Polymerase.
PCR Reaction
25 µL [TAPS-HCl, pH9.3, 50 mmol/l KCl, 2 mmol/l MgCl2, 1 mmol/l 2-Mercaptoethanol, 0.01 % gelatin, 200 µmol/l dNTPs, 0.2 M primer, 1.25 units Taq DNA Polymerase]
Lane 1 : Ethachinmate 0 µl
Lane 2 : Ethachinmate 0.2 µl
Lane 3 : Ethachinmate 0.5 µl
Lane 4 : Ethachinmate 1 µl
Lane 5 : Ethachinmate 3 µl
Lane 6 : Ethachinmate 5 µl
Result
Ethachinmate did not affect the activity of Taq DNA Polymerase.
