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Volume 20, Number 5—May 2014
Research

Bovine Leukemia Virus DNA in Human Breast Tissue

Gertrude Case BuehringComments to Author , Hua Min Shen, Hanne M. Jensen, K. Yeon Choi1, Dejun Sun, and Gerard Nuovo
Author affiliations: University of California, Berkeley, California, USA (G.C. Buehring, H.M. Shen, K.Y. Choi, D. Sun); University of California Davis Medical Center, Sacramento, California, USA (H.M. Jensen); Ohio State University Comprehensive Cancer Center, Columbus, Ohio, USA (G. Nuovo)

Main Article

Table 4

Primers and reaction conditions for GAPDH gene amplifications used to verify quality of BLV DNA extracted from human breast tissue samples, human cell lines, and animal cell lines*

Gene Primer pair sequences, 5′ → 3′† Location in bp‡ Product length, bp Annealing temperature, °C§ Extension time, s§
Human GAPDH F: GAGTCAACGGATTTGGTCGT 194–213 237 50 22
R: TTGATTTTGGAGGGATCTCG 431–412
Mouse GAPDH F: AGCTTGTCATCAACGGGAAG 246–265 796 58 60
R: ATGTAGGCCATGAGGTCCAC 1041–1022
Bovine GAPDH F: CCTTCATTGACCTTCACTACATGGTCTA 172–199 857 59 60
R: GCTGTAGCCAAATTCATTGTCGTACCA 1028–1002

*GAPDH, glyceraldehyde-3-phosphate dehydrogenase; BLV, bovine leukemia virus; F, forward primer; R, reverse primer.
†Reverse sequences are reversed and complementary to the published genomic sequences. Primers were synthesized by Operon Biotechnologies, Huntsville, AL, USA.
‡Sequence bp numbering according to GenBank no. NM 002046.4 (human), XM 001476707.3 (mouse), and NM 001034034.2 (bovine).
§Cycling conditions were 1 cycle at 95°C for 2 min; 35 cycles at 95°C for 30 s; XX°C for 30 s, 72°C for XX sec; and 1 cycle at 72°C for 5 min, where XX represents annealing temperature and extension times, respectively, for primer pairs.

Main Article

1Current affiliation: Texas A&M Health Science Center College of Medicine, College Station, Texas, USA.

Page created: April 16, 2014
Page updated: April 16, 2014
Page reviewed: April 16, 2014
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