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#16S rDNA Sanger  Sequencingand Analysis (Organism Identification)  Following liquid culturing, the organisms need to be identified, or classified. This is accomplished by determining and then analyzing the DNA sequence of the 16S rRNA gene. In this section, we describe how the sequence of this gene is determined and readied for analysis. The general outline is as follows: DNA extraction, polymerase chain reaction (PCR) amplification of the 16S rRNA gene, and sequencing of the resulting PCR product using Sanger sequencing \cite{Sanger_1977}. There are multiple approaches one can take to these steps. For example, the PCR requires DNA from the organisms of interest. That DNA can come directly from a liquid culture of the organism (when this is used for PCR this is known as colony PCR). Alternatively, one can take a liquid culture and then isolate the DNA from that culture and use the purified DNA as material for the PCR. This adds an extra step to the process - a step known as DNA extraction (see below.) Colony PCR significantly decreases the amount of work needed for preparation, but it can yield poorer results, both in terms of PCR success and resultant sequence quality. However, we recommend colony PCR when screening a large number of samples. DNA extraction can then be used for any recalcitrant samples. DNA extraction is significantly more work, but it often generates better Sanger sequences allowing for more accurate identification.  ##DNA Extraction 
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SeqTrace provides excellent directions for using the program at [https://code.google.com/p/seqtrace/wiki/WorkingWithProjects](https://code.google.com/p/seqtrace/wiki/WorkingWithProjects)  ##Edit ###Edit  and Create a Consensus Sequence with SeqTrace For this workflow we have found that the following is the simplest way to edit and create a consensus sequence from a forward and reverse read in SeqTrace.   1. Create a new project (File > New Project)