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Rapid amplification of cDNA ends
The protocols for 5′ or 3′ RACES differ slightly. 5′ RACE-PCR begins using mRNA as a template for a first round of cDNA synthesis (or reverse transcription) reaction using an anti-sense (reverse) oligonucleotide primer that recognizes a known sequence in the middle of the gene of interest; the primer is called a gene specific primer (GSP). The primer binds to the mRNA, and the enzyme reverse transcriptase adds base pairs to the 3′ end of the primer to generate a specific single-stranded cDNA product; this is the reverse complement of the mRNA. Following cDNA synthesis, the enzyme terminal deoxynucleotidyl transferase (TdT) is used to add a string of identical nucleotides, known as a homopolymeric tail, to the 3′ end of the cDNA. (There are some other ways to add the 3′-terminal sequence for the first strand of the de novo cDNA synthesis which are much more efficient than homopolymeric tailing, but the sense of the method remains the same).
PCR is then carried out, which uses a second anti-sense gene specific primer (GSP2) that binds to the known sequence, and a sense (forward) universal primer (UP) that binds the homopolymeric tail added to the 3′ ends of the cDNAs to amplify a cDNA product from the 5′ end.
Rapids are hydrological features between a run (a smoothly flowing part of a stream) and a cascade. Rapids are characterised by the river becoming shallower with some rocks exposed above the flow surface. As flowing water splashes over and around the rocks, air bubbles become mixed in with it and portions of the surface acquire a white colour, forming what is called “whitewater”. Rapids occur where the bed material is highly resistant to the erosive power of the stream in comparison with the bed downstream of the rapids. Very young streams flowing across solid rock may be rapids for much of their length. Rapids cause water aeration of the stream or river resulting in better water quality.
Smashes a single target with the equipped shield five times, where each hit will inflict melee physical damage. The damage is affected by the equiped shield’s weight and upgrade level, and its hit rate is increased by 20.