An important problem in measurement of messenger RNA (mRNA) levels by reverse transcription-polymerase chain reaction (RT-PCR) is DNA contamination, which can produce artifactually increased mRNA concentration. Current methods to eliminate contaminating DNA can compromise the integrity of the RNA, are ...
RNA contaminationcarcinogen‐DNA adduct32P‐postlabelingDNA isolation32P-Postlabeling is a widely applied assay for the analysis of carcinogen-DNA adducts. Optimization of most steps in this assay has been given attention, but influences of DNA isolation and DNA purity on adduct quantitation have not...
contamination is still present. Centrifuge the precipitate, resuspend the pellet in nuclease-free water, and extract with phenol:chloroform:IAA until this quick-forming white precipitate no longer occurs. High yields of intact RNA can be achieved once abundant DNA and ...
Panel A shows high integrity and quality of isolated RNA; Panel B shows the RNA linear recovery; Panel C shows no cross contamination and efficiency in DNA removal. Panel D shows its application in siRNA evaluation (Human umbilical vein endothelial cells were transfected and mRN...
This was examined with a view that a positive mtRNA rate would indicate low contamination. To ensure that reads originating from 'Nuclear mitochondrial DNA sequences' (NUMTs) were not mistaken for mtRNA mapping reads, we only considered uniquely mapping reads in STAR. The rRNA rate was defined...
三、污染与阴性对照(Contamination and No-template Controls) 1. 进行qPCR实验时,应保证良好的实验操作规范,以防对工作区域、试剂、耗材、仪器、DNA标准品等造成污染。推荐将反应体系配制区和模板制备区进行物理隔离,并定时使用0.5%次氯酸钠或10%漂白剂对各实验区域进行擦拭清理。
Additional sample amplification steps are frequently required, leading to prolonged experimental durations (~2 h) and significantly increasing the risk of aerosol contamination. Although other CRISPR enzymes, such as CRISPR-Cas13a, enable direct detection of miRNA, they still pose challenges in ...
3. Add 1 ml TRIzol to the plate to lyse the cells. Pass the cell lysate several times through a pipette (insufficient TRIzol may result in contamination of the isolated RNA with DNA). 4. Transfer the lysate to a 1.5 or 2.0ml Eppendorf tube. ...
Therefore, if the A260 of the sample is higher than expected and protein and phenol contamination have been ruled out, the sample is likely contaminated with DNA. In this case, DNase treatment followed by phenol extraction and ethanol precipitation is recommended (see Purification of RNA by SDS...
It is specific—performed measurement is selective for DNA or RNA It is sensitive—can measure pg/mL; it is the recommended method for very diluted nucleic acid samples It is accurate despite contamination being present in the sample, including nucleic acid contaminants What are the disadvantages?