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The use of real-time PCR is a robust and sensitive technique. However, it is essential to ensure that there are no inhibitors in the nucleic acid extract. If inhibitors were present, then quantification of the gene of interest could not be achieved. The use of real-time PCR is therefore only suited to samples where inhibitors are well controlled in the purified DNA. In this study, a standard curve was developed for use of the TaqMan PCR assay for the determination of the number of Actin-eubacterial cells in sediment samples. A standard curve was created from serial dilutions of eubacterial cells, which was determined to be linear over 8 orders of magnitude. The detection limit was determined to be 1.5 × 106 cells mL-1. The sensitivity and reproducibility of this assay was determined by assessing the linearity, reproducibility and amplification efficiencies of the assay. The assay was linear from 0.005 to 50 cells mL-1 for the minimum threshold cycle (Ct) and the efficiency of the amplification was 100% over the 5-log range. The real-time PCR assay was used to determine the total eubacterial number in sediment samples. The assay was linear over a 4-log range and the efficiency of amplification was 100%. This study is the first to describe the use of real-time PCR to amplify a housekeeping gene from environmental DNA. This study also describes the use of real-time PCR to measure bacterial numbers in sediment samples. Copyright © 2018 Elsevier B.V. All rights reserved.

npm debug % is not working. It says i dont have the game installed. Its an application to play Sims 3. Please help me install this so i can play game and not have to waste money on the game. Please help

Answer: You can do so by simply re-installing a fresh copy of the game. For all other games, you do not have to re-install the game. So, you just need to download a fresh game. So, we recommend you to download a fresh copy.

Hi, I downloaded the game but when I was trying to install it it was asking to download the expansion from steam and it wouldn't allow me to and it keeps on crashing i don't know what to do or what i did wrong. Please help

How do I install this on a steam client. I have the game, but when I was tryiong to install it I had to download the expansion, but when I did it crashed my laptop. Please help me, I really need this game! 827ec27edc