How does DNA polymerase distinguish between rNTP & dNTP? What are the differences and similarities between cDNA and mRNA? Why is cDNA used as a template in RT-PCR, not mRNA? What is the difference between the purpose of DNA cloning and ...
Molecular difference between WASP and N-WASP critical for chemotaxis of T-cells towards SDF-1α Neeraj Jain & Thirumaran Thanabalu Wiskott-Aldrich Syndrome protein (WASP) integrates cell signaling pathways to the actin cytoskeleton, which play a critical role in T-cell activation and ...
2.2. RNA isolation and cDNA synthesis Total RNA was isolated and cDNA was prepared using the RNeasy minikit (Qiagen) and SMART PCR synthesis kit (Clontech Laboratories), respectively, according to manufacturer's instructions. First-strand cDNA synthesis was performed with reverse transcriptase (RT M...
Transcripts were quantified using a combination of competitive reverse transcriptase PCR (RT-PCR) and t-RFLP. In addition to pmoA, we used mmoX encoding for a subunit of the sMMO to search for genes and transcripts of aerobic MOB that lack a pmoA. Three different primer sets were applied ...
RT-qPCRMicroRNA-21 (miR-21) overexpression is characteristic for various types of tumors, but it is still unknown whether its expression levels differ between invasive and non-invasive breast carcinomas. The main goal of the study was to determine the difference in miR-21 expression among ...
and transfer to the target cells. Upon receiving a signal by the receptors, the concentration of these molecules in the cell increases and leads to the alteration of one or moreenzymesin the cell. The key difference between cAMP and cGMP is thatcAMP is synthesized from theATPby adenylyl ...
Bloodstain age can be determined by measuring the 18S rRNA:尾-actin mRNA ratio by Reverse Transcription-quantitative PCR (RT-qPCR). Since this ratio is a function of time, it can be used as an estimator of bloodstain age. However, it is important to validate the technique in a variety ...
2.7. Reverse Transcription-Quantitative PCR (RT-qPCR) The mRNA levels of selected genes were analyzed by RT-qPCR. For mRNA expression in mouse RPE, total RNA was prepared by the two-step extraction method as described above. First-strand cDNA was synthesized from 200 ng of total RNA with ra...
2.8. Isolation of Total RNA and qPCR The isolation of total RNA was performed using the NucleoSpin RNA XS Kit (Macherey-Nagel GmbH & Co. KG, Düren, Germany). First-strand complementary DNA (cDNA) synthesis was performed using 650 ng of total RNA as previously described [31]. Real-time...
2.4. Fluorescence Quantitative PCR (qPCR) The DEGs involved in carotenoid metabolism synthesis were screened and validated by qPCR. RNA was reverse transcribed using the TaKaRa PrimeScriptTMRT reagent kit. Primers were designed using Primer 3, andRPS18(FJ608659) andα-TUB(FJ526336) were the inte...