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Site-directed mutagenesis primers (Table 2) were designed to replace the relevant amino acid residue with alanine and carried out using the PfuUltra II Fusion HS DNA Polymerase kit (600670, Agilent) according to the manufacturers instructions

There is also a third pathway that makes nasal delivery uniquely interesting for certain applications

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2 Common symptoms include: Weight gain and difficulty losing weight Fatigue and low energy Feeling cold Dry skin and hair Constipation Depression and low mood Brain fog and poor concentration Muscle aches and weakness Hypothyroidism is diagnosed with a blood test measuring TSH (a hormone that tells our thyroid how much to produce)