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A High Resolution Melting (HRM) assay (Biomarkers, Carlsbad, CA) and LightCycler 480 System (Roche, Indianapolis, IN) were used to screen subjects for point mutations in *APC*. Briefly, PCR products were subjected to two separate denaturation steps, one at 95°C for 5 minutes and the other at 94°C for 30 seconds, followed by an annealing step at 65°C for 20 seconds. The HRM reactions were performed at 94°C for 5 minutes, followed by 37°C for 15 minutes, and then held at 40°C for 1 minute. Melting curves were generated from 65°C to 95°C at 0.01°C/sec. For screen-positive individuals, the PCR products were sequenced by the personnel at Mass Spectrometry Analysis Laboratory of the University of Nebraska Medical Center (Omaha, NE).

Molecular Analysis of *APC* {#s4_2}
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*APC* exon 15 was amplified from genomic DNA of peripheral blood lymphocytes by PCR using primers APC15.1 and APC15.2, and fragment length was confirmed by agarose gel electrophoresis. Sequences of PCR products were determined by the personnel at DNA Analysis Laboratory of the University of Nebraska Medical Center.

*APC* exon 15 mutations were classified into three groups based on their frequencies in the Czech population (Table [1](#T1){ref-type=”table”}). The first group included mutations with a frequency higher than 6%. Other mutations were classified as rare. In the rare group, mutations with a frequency of less than 2% were categorized as rare, while mutations with a frequency of 2% to 4% were categorized as polymorphic (Table [1](#T1){ref-type=”table”}). Point mutations and deletion/insertion mutations in *APC* were identified using the online databases UNIPROT (), Mutation Taster (), and PolyPhen2 (

Rainbow Six: Vegas 2 is the sequel to the 2010 tactical shooter Rainbow Six: Vegas. After the events of the first game in the series the player will be given a full control over six operators of the covert military unit “Grey Ghost”.

Rainbow Six: Vegas 2 is an action game set in Las Vegas where the first strike of the terrorist attack has taken place. The players will have to infiltrate an Arab terrorist group and foil their plans. You will have to use the abilities of the special operatives of the Rainbow Team to infiltrate buildings in the city and defeat terrorists.

Rainbow Six: Vegas 2 features six operatives that will be available to you. Each team member will have his or her own abilities, weapons and gadgets that will be used in the game.

Rainbow Six: Vegas 2 is a turn-based tactical shooter that offers you unprecedented level of freedom and the ability to take advantage of the situations using a variety of authentic and improved weapons and gadgets.

The Rainbow Six: Vegas 2 key features include:

6 operatives, each with their own individuality;

A vast selection of guns, including the 70mm Claymore shotgun, a new semi-automatic pistol, a silenced pistol with an extended magazine, a suppressed pistol that fires lasers, a shotgun with a flash bang, a Taser, grenades and others;

A wide range of abilities;

The best intel support of the previous games;

Easily control your operatives in three different levels of difficulty: green light, orange light and red light;

Over a dozen of city locations;

Improved multiplayer gameplay;

40 challenging missions set in the most exciting, exciting places in the world.

Rainbow Six: Vegas 2 also has the Rainbow Six Dynamic Open World experience: each and every city has its own layout and story, and every map and location is created by the player using a variety of unlocks and achievements.

Rainbow Six: Vegas 2 is a tactical shooter that requires strategic and accurate thinking for each situation.

In terms of gameplay, each level is divided into two halves: team play and freeplay. The freeplay is the element of the game when it is not allowed to progress through the levels using the abilities of the Rainbow Team members.

In team play mode the players can progress through the levels on their own, and they can use their own abilities, weapons and gadgets in order to counter the enemy and complete the mission.

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