생체자기비즈 전문제조업체
결과를 개선하는 방법 260/230 시약추출을 위해 자기비드법을 이용한 계산?
Magnetic bead 방식을 이용하여 핵산을 추출하는 경우, 낮은 260/230 비율 (대개 < 1.8) 유기 화합물의 존재를 나타냅니다.(구아니딘 염과 같은, 페놀, 에탄올, EDTA) or residual salts in the sample, which can affect downstream experiments (PCR과 같은, 시퀀싱).
Here is a systematic optimization plan:
- Optimization of Extraction Steps
에이. Complete sample lysis
Problem: Incomplete lysis leads to incomplete release of impurities.
Improvement: Increase the lysis time (예를 들어, extend to 30 분) or temperature (56℃).
Add a grinding step or additional proteinase K digestion for complex samples (such as plants, 대변).
비. Thoroughly wash the magnetic beads
Key step: Incomplete washing is the main cause of 260/230 낮은.
Improvement: Increase the number of washes: 2 times → 3 타임스 (especially for samples with high impurities).
Washing solution formula: Ensure to use freshly prepared 80% 에탄올 (avoid excessive water content or contamination).
Washing operation: Vortex or shake to mix the magnetic beads each time, avoiding the formation of beads clumps.
Thoroughly aspirate the supernatant (can briefly centrifuge to remove residual liquid).
기음. Complete drying of magnetic beads
Problem: Residual ethanol can significantly reduce 260/230.
Improvement: Open the cap and let it dry for 5-10 분 (at room temperature or 37°C), until there is no ethanol smell. Avoid excessive drying (otherwise, it will reduce the efficiency of nucleic acid elution).
d. Optimize elution conditions
Use preheated elution buffer (such as 65°C TE buffer or sterile water), and let it stand for 5 minutes before collection.
Avoid using too small elution volume (recommend ≥ 50 μL to reduce salt concentration).
Ⅱ.품질 관리 of Reagents and Consumables
에이. Replace with fresh reagents:
Check if ethanol and washing solution have been contaminated (if stored for a long time without sealing).
Avoid using expired magnetic bead reagents (magnetic bead degradation may adsorb impurities).
비. Consumables selection:
Use centrifuge tubes and pipettes without nuclease or pyrogen (to avoid interference from plastic additives).
Ⅲ.Sample preprocessing
- Complex samples (such as soil and plants):
Increase the CTAB or SDS pretreatment to remove polysaccharides and polyphenols.
- For blood samples:
First remove hemoglobin using erythrocyte lysis solution.
- Inhibitor-enriched samples (such as feces and body fluids):
Dilute the samples or use dedicated lysis buffers (such as those containing Carrier RNA).
Ⅳ.Verification and remedial measures
에이. Electrophoresis or Qubit verification:
Check for degradation (tailing phenomenon) using agarose gel electrophoresis.
Compare the results with those from Nanodrop using a fluorescence meter (Qubit) to eliminate interference from impurity absorbance.
비. Purification remediation:
If the ratio of 260/230 remains low, purify the nucleic acid using a silica column or by ethanol precipitation again.
다섯. Control Experiment
For each extraction, a known standard sample (such as λDNA or cultured cells) is set up to confirm the stability of the reagents.
If the 260/230 ratio of the control sample is normal, the problem may lie in the sample being tested itself.
Common Problem Troubleshooting Checklist
| Phenomenon | Possible Cause | Solution |
| 260/230 is low but 260/280 is normal | Protein or phenol contamination | Strengthen the washing and drying steps |
| Both 260/230 그리고 260/280 are low | Protein or phenol contamination | Optimize the lysis, replace the lysis solution |
| Only the 230 peak is high | Organic solvent contamination | Check the purity of ethanol or buffer |
By following these steps, the ratio of 260/230 can be significantly increased (target > 1.8). If the problem persists, please feel free to contact Shanghai Lingjun Biotechnology at 18103799069.
공급자
상하이 링쥔 생명공학 유한회사, 주식회사에 설립되었습니다 2016 생체자성재료 및 핵산추출시약 전문제조업체입니다..
핵산 추출 및 정제 분야에서 풍부한 경험을 보유하고 있습니다., 단백질 정제, 세포 분리, 화학발광, 및 기타 기술 분야.
우리의 제품은 많은 분야에서 널리 사용됩니다., 의료 테스트와 같은, 유전자 검사, 대학 연구, 유전자 육종, 등. 우리는 제품을 제공할 뿐만 아니라 OEM도 수행할 수 있습니다., ODM, 그리고 다른 필요. 관련된 필요사항이 있는 경우, 저희에게 연락하게 자유롭게 느끼십시오 .
