影响pcr反应的因素: 1.模板纯度,有较多蛋白或其他杂质时,会一定程度影响pcr效率; 2.模板量,过高的模板量反而会降低pcr效率和特异性; 3.引物,引物的长度需要再效率和特异性间获得优化,其次要控制引物gc含量; 4.聚合酶,要考虑酶的保真率和速度; 5.mg离子浓度,mg离子浓度过高会降低特异性,浓度过低会降低效率; ...
The blocking buffer was aspirated from the protein A-coated plate, and the chromatin + antibody mix from the untreated plate was transferred to the protein A-coated wells and incubated in the ultrasonic bath for 60 min at 4°C. The wells were then washed 3 times with immunoprecipitation ...
N6-methyladenosine (m6A) is the most abundant posttranscriptional modification of mRNA in eukaryotes. Recent evidence suggests that dysregulated m6A-associated proteins and m6A modifications play a pivotal role in the initiation and progression of diseas
Tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral ribonucleic acid (RNA) using reverse transcription polymerase chain reaction (RT-PCR) are pivotal to detecting current coronavirus disease (COVID-19) and duration of detectable virus indicating potential for infectivity. Metho...
Zip1+/+, Zip1+/− and Zip1−/− mice were identified using two pairs of PCR primers (Primer pair 1: F1: 5’-TCTCTGTATAGGTCCAAGAGTCAC-3’, R1: 5’-TGAAGGAGGGACAAGCAAATA. Primer pair 2: F2: 5’-CATGTGACGGTAAGCATTGACTTC-3’, R1: 5’-TGAAGGAGGGACAAGCAAATAAGTG-3’). ...
Plasma samples were taken from 348 patients, aged 18 to 85 years, who were 45.11% female (n= 157) and 54.89% male (n= 191). The stage of COVID-19 infection varied from day 1 of clinical manifestation (and/or first positive PCR result) to 438 days since an officially registered COVI...
Quantitative PCR analysis of these genes (Figure S5) showed a significant increase in ACE and AGT at 24 h, whereas renin was drastically downregulated (more than 50% reduction) at the same time. As with the previous data, neither of the aforementioned genes was significantly modulated at 48 ...
Each amplification reaction was performed using 100 ng (2 μL) of genomic DNA in 25 μL of reaction mixture consisting of 1.0 μL of each primer (10 μmo1/L), 12.5 μL 2 × Taq PCRMasterMix (constituent: 0.1 U Taq polymerase/μL, 500 μM dNTP each, 20 mM Tris-HCl, pH 8.3,...
1c). When methylated or silenced HNE1 and HONE1 nasopharyngeal cells were treated with the demethylation reagent 5-aza-2′-deoxycytidine (Aza) with or without trichostatin A (TSA) histone deacetylase, PCR confirmed that TET1 expression was restored with a significant decrease of TET1 promoter ...
(Promega C1181) in a final volume of 20 μl, following the manufacturer’s protocol. RT minus controls are obtained by diluting the same volume of RNA in 20 μl of DEPC treated water. 4 μl of cDNA were used in the PCR reaction (GoTaq DNA polymerase, Promega M300) in a final ...