How to Choose the Right PCR ReagentsTanuja Koppal, Ph.D
PCR The polymerase chain reaction (PCR) is a fast and inexpensive technique for amplifying a DNA sequence of interest. It consists of three steps: Denaturation: The sample is heated to separate the DNA into two single strands. Annealing: The temperature is lowered to allow primers to anneal...
Explain the process of PCR and the reagents that are needed to perform the reaction. Why can't DNA polymerase initiate the synthesis of a new DNA strand? How did the discovery of the enzyme DNA polymerase play in the understanding for the process of DNA replication?
Clean-up of PCR reaction Once the template DNA has been amplified by PCR, clean-up is required to remove unincorporated primers; any leftover primers can interfere with the strand-specific cycle sequencing reactions that will follow. There are several ...
After adding the nucleic acid to be detected, the reaction system performs a fluorescent PCR amplification reaction. The time is slightly different depending on the PCR reagents and instruments used. Generally,it takes 90-120 mi...
These DNA substrates were consequently introduced into a human cell line (HCT 116), and joining was characterized both in terms of efficiency, by quantitative PCR (qPCR) of head-to-tail products, as well as product structure, by sequencing (Fig. 2a and Supplementary Fig. 2). We used ...
When real-time polymerase chain reaction came out during the 90s the words kinetic PCR analysis or real-time PCR were most commonly used [1][2][3]. Also, real-time quantitative RT-PCR was used to indicate RT-qPCR [4]. When real-time PCR became more common in the 2000s it was more...
This method has certain inherent limita- tions, including long reaction times and leakage of light from one microarray feature to its neighbors, both of which introduce unwanted changes into probe sequences. While microarrays with probes synthesized in this way can generate reproducible and ...
Manual qPCR set-up requires significant knowledge and skill, and operators must take precautions to avoid cross-contamination. Despite this, operator error remains the most likely cause of qPCR contamination. qPCR reaction contamination is a serious problem, as nucleic acid contamination can prod...
Heterodimeric capping protein (CP/CapZ) is an essential factor for the assembly of branched actin networks, which push against cellular membranes to drive a large variety of cellular processes. Aside from terminating filament growth, CP potentiates the n