MB ASCP Study Aids Buckingham Ch. 7 2023 Updated.

MB ASCP Study Aids Buckingham Ch. 7 2023 Updated.Polymerase Chain Reaction (PCR) - ANSWERPrimer-directed DNA polymerization in vitro resulting in amplification of specific DNA sequences. Target amplification - ANSWERTarget: Specific sequences or regions that are detected in an analytical procedure. Amplification: Replication/copying Target amplification: replication of a specific target region of DNA. Template - ANSWERSingle stranded nucleic acid used as a guide for replication or synthesis of a complementary strand. Deoxynucleotide triphosphates (dNTP) - ANSWERdNTP, (dATP, dGTP, dCTP, dTTP) Strand - ANSWEROne molecule of nucleic acid. A double helix contains two strands of DNA. Amplicons - ANSWERProduct of a PCR reaction. Copies of DNA. Components of a PCR reaction - ANSWEROligodeoxynucleotide primers, dNTPs, KCl, buffer, MgCl2, Polymerase, Template 3 steps of PCR - ANSWERDenaturation, Annealing, Extension Denaturation Step - ANSWEROne double stranded DNA target is denatured into two single strands by heating the sample 94-96 degrees C. Annealing Step - ANSWERTwo oligonucleotide primers anneal (hybridize) to complementary sequences on the template. Temperature ranges from 50-70 degrees C, optimized with specific primer. Extension Step - ANSWERPolymerase synthesizes a copy of the template DNA by adding nucleotides to the hybridized primers. Temperature occurs from 68-72 degrees C, the optimal temperature of the enzyme. Mispriming - ANSWERAberrant initiation of DNA synthesis from a primer hybridized to template sequences different from the intended target.