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This contrasts with acetyl-L-carnitine (CAS: 14992-62-2, molecular formula: C9H17NO4, molecular weight: 203.24 g/mol), which appears as a white crystalline powder, providing two-carbon acetyl ester precursors for acetyl-CoA through acetylation of L-carnitine with acetic anhydride or enzymatic synthesis

When we included LiPA results for those samples in which sequencing was not done, overall agreement at the subtype level (with either LiPA or sequencing) even slightly increased to 98.6% (140/142), as the agreement at HCV-1 subtype calling between the GT Plus and the LiPA assays was 100% (all being 1b except one 1a case)

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PCR was performed with a 2 l cDNA mixture in a total volume of 35 l using Taq polymerase and 0.2 nmol/l of each forward and reverse primer (intron spanning, 5CCCCCGATAGCTCCTACAAC3 and 3AAGGCCTTCCTCATGCACT5, TRPV1 mRNA accession number NM_018727) after initial 95 C for 3 min over 35 cycles of 95 C for 30 s, 64 C for 30 s and 72 C for 45 s, finalized by 72 C for 7 min and a temperature hold at 4 C in a Techne TC-512 gradient thermal cycler (Staffordshire, UK), yielding a 295 bp product

The cells were incubated at 37 C under a 5% CO 2 /95% air atmosphere