PCR (Google e-Livro)
A thoroughly updated version of the successful first edition with a new chapter on Real-Time PCR, more prokaryotic applications, and more detail in the complex mutagenesis sections. Information on PCR applications in genomics and proteomics have been expanded and integrated throughout the text. There is also advice on available products and specific pointers to the most appropriate methods. As with the first edition, this will be an ideal practical introduction and invaluable guide to PCR and its applications.
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1 An introduction to PCR
2 Understanding PCR
3 Reagents and instrumentation
4 Optimization of PCR
5 Analysis sequencing and in vitro expression of PCR products
6 Purification and cloning of PCR products
7 PCR mutagenesis
adaptor addition agarose gel aliquot alleles allow amino acid amplicon ampliﬁcation products amplify analysis annealing temperature approach appropriate assay base pairs buffer cells Chapter cloning codon coli complementary concentration contain deﬁned denaturation detection difﬁcult DNA fragments DNA sequence DNA synthesis dNTPs double-stranded efﬁcient ensure example exonuclease ﬁdelity Figure ﬁnal ﬁrst ﬁrst-strand cDNA ﬂanking gel electrophoresis genomic genomic DNA hybridization identiﬁed increase incubation insertion labeled ligation microcentrifuge modiﬁed molecular beacons mRNA mutagenesis mutations nested PCR nonspeciﬁc Northern blot Nucleic Acids nucleotides oligo-dT oligonucleotide oligonucleotide primers optimization PCR ampliﬁcation PCR cycle PCR primers PCR products PCR reaction performed plasmid polymorphisms primers primers P1 probe proofreading protein Protocol puriﬁcation reagents real-time RT-PCR recombinant region restriction digestion restriction enzyme reverse transcriptase RT-PCR samples single-stranded speciﬁc standard step strand sufﬁcient Taq DNA polymerase TaqMan target gene template DNA thermal cycler thermostable DNA polymerase transcription tube vector vectorette wild-type