Sequence analysis bottom

Fig. 6. DNA sequence analysis, (a) Simplified methodology for dideoxy sequencing. A primer, 5 -TCTA, hybridized to the template, is used to initiate synthesis by DNA polymerase, (b) Stmcture of 2, 3 -dideoxy CTP. When no 3 -OH functionaUty is available to support addition of another nucleotide to the growing chain, synthesis terminates once this residue is incorporated into the synthetic reaction, (c) Representation of a DNA sequencing gel and the sequence, read from bottom to the top of the gel, gives sequence information in the conventional 5 to 3 direction.

From Jornvall and Pietruszko (73) for sequence analysis and from Berger et al. (7S) for total compositions of tryptic peptides. In the latter case residues have been ordered to yield maximum homology with the other structures. Peptides were similarity positioned in both studies. Empty spaces not analyzed. Known subunit differences are the bottom line alternatives at position 43 (73) affecting one set of subunits, and at 230 (7S) affecting another set (the atypical enzyme). 

Luckas, W. J. et al., A Human Reliability Analysis for the ATWS Accident Sequence at the Peach Bottom Atomic Power Station, BNL Technical Report A3272, May 1986. 

Operational sequence diagrams are flcw-charting techniques that represent any sequence of control movements and information collection activities that are executed in order to perform a task. Various activities in the diagram are represented with a symbolic notation, supported where necessary by a text description. For the majority of simple applications, OSDs assume a linear flow drawn from top to bottom with a limited degree of branching and looping. The symbols used are usually tailored to fit the type of task being studied and its level of analysis. 

The second and most important finding of this sequencing project was that the difference of 14 Da between the two proteins was not because of a methy-lation of the protein isolated from chicken but because of a substitution of valine for leucine. The MS/MS analysis of the tryptic pieces in which the substitution occurred is shown in Figure 10.11. Had this been a bottom-up... 

In recent years, a novel approach to protein identification emerged, called top-down sequencing. Here the entire nondigested protein is analyzed. Apart from accurate MW measurement, the protein ion is fragmented by the electron capture dissociation (ECD) method (see Chapter 3). This provides in-depth information on the sequence of protein. Such analysis can be performed only with FTICR instruments (see Section 2.2.6) that ensure high resolution and accuracy but, at the same time, they are exceptionally expensive. However, as very large ions are analyzed, even the high accuracy of FTICR is sometimes not sufficient, and it is recommended that such analyses are accompanied by more traditional bottom-up approaches. 

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