Instead of promoting irreversible primer extension like the Sanger method, the reversible chain terminators method uses a cyclic method that consists of nucleotide incorporation, fluorescence imaging and cleavage. The figure below shows a modified nucleotide with a cleavable dye and reversible blocking group. Once the blocking group is removed, a new nucleotide may come in.
The steps for such a process can be outlined as follows:
Have four dNTP's, each with a different fluorescent marking. These markings should not interfere with base pairing or phosphodiester bond formation.
Each dNTP should terminate DNA elongation temporarily with a blocking group on the 3' carbon of the sugar moiety.
Upon each cycle, have just one dNTP bind to the elongating strand and emit a fluorescent dye color.
Depending on the color emitted, record the particular nucleotide.
Cleave the blocking group and fluorescent dye with a palladium-catalyst.
Restore a 3' hydroxyl so that the growing strand can now elongate.
Repeat from step 1.
There are some limitations to this method which include:
Linux for Beginners doesn't make any assumptions about your background or knowledge of Linux. You need no prior knowledge to benefit from this book. You will be guided step by step using a logical and systematic approach. As new concepts, commands, or jargon are encountered they are explained in plain language, making it easy for anyone to understand.
This book is designed to be used as the primary textbook in a college-level first course in computing. It takes a fairly traditional approach, emphasizing problem solving, design, and programming as the core skills of computer science. However, these ideas are illustrated using a non-traditional language, namely Python.