Genotyping of Methylenetetrahydrofolate Reductase Gene by Pyrosequencing Coupled with Polymerase Chain Reaction Using Human Whole Blood as Starting Material

2013 ◽  
Vol 40 (7) ◽  
pp. 1037-1042
Author(s):  
Yun-Long LIU ◽  
Zhi-Yao CHEN ◽  
Hai-Ping WU ◽  
Guo-Hua ZHOU
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Methylenetetrahydrofolate Reductase ◽  
Starting Material ◽  
Chain Reaction ◽  
Human Whole Blood ◽  
Reductase Gene ◽  
Polymerase Chain ◽  
Methylenetetrahydrofolate Reductase Gene

scholarly journals Structurally and functionally conserved regions of cytochrome P-450 reductase as targets for DNA amplification by the polymerase chain reaction. Cloning and nucleotide sequence of the Schizosaccharomyces pombe cDNA

1992 ◽  
Vol 287 (1) ◽  
pp. 195-200 ◽  
Author(s):  
J S Miles
Keyword(s):  
Polymerase Chain Reaction ◽  
Schizosaccharomyces Pombe ◽  
Binding Domain ◽  
Amino Acid Sequences ◽  
Chain Reaction ◽  
Conserved Regions ◽  
Reductase Gene ◽  
Degenerate Oligonucleotide ◽  
Polymerase Chain ◽  
Cytochrome P 450

1. Alignments of the available cytochrome P-450 reductase amino acid sequences, and comparison with the crystal structure of ferredoxin-NADP reductase, indicate that two highly conserved regions are of functional importance. 2. Degenerate oligonucleotide primers, based on these sequences, were used in the polymerase chain reaction to amplify a 309 bp fragment of the cytochrome P-450 reductase gene from Schizosaccharomyces pombe for use as an homologous probe. 3. A 2.6 kb cDNA was cloned from a lambda library, and sequencing revealed an open-reading frame of 2034 bp encoding a protein of M(r) 76774. This protein shares 38-41% identity with other eukaryotic cytochrome P-450 reductases, and 30% identity with that of Bacillus megaterium. 4. Comparison of the N-terminal FMN-binding domain with flavodoxin, and the C-terminal FAD- and NADP-binding domain with ferredoxin-NADP reductase, indicates the presence of several functionally conserved regions. 5. The Sc. pombe cytochrome P-450 reductase gene was shown to contain no introns.

A novel buffer system, AnyDirect, can improve polymerase chain reaction from whole blood without DNA isolation

2007 ◽  
Vol 380 (1-2) ◽  
pp. 112-117 ◽  
Author(s):  
Young Geun Yang ◽  
Jong Yeol Kim ◽  
Young-Han Song ◽  
Doo-Sik Kim
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Dna Isolation ◽  
Buffer System ◽  
Chain Reaction ◽  
Polymerase Chain
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