Pseudogene-Free Amplification of Human GAPDH cDNA

Abstract

One apparent pitfall of the technique was the fact that reactions were carried out first on the plate supernatants and then on individual plugs from the plates, suggesting the possibility of a general contamination of the plugs. We could see that it was not the case and that the contamination level was very low compared with the signal obtained with positive plugs. Therefore, this technique makes it possible to screen a phage library in about a week, without the expense of membranes, and at a high stringency using PCR. REFERENCES 1.Israel, D.I. 1993. A PCR-based method for high stringency screening of DNA libraries. Nucleic Acids Res.

Chemokine (C-C Motif) Receptor 1 Is Required for Efficient Recruitment of Neutrophils during Respiratory Infection with Modified Vaccinia Virus Ankara

“…and converted to cDNA using Omniscript reverse transcriptase (Qiagen). PCR was carried out as described previously (25). Primer pairs for amplification of murine GAPDH (glyceraldehyde-3-phosphate dehydrogenase), specific murine chemokines, and the VACV E3L gene were designed using Primer3 software (26).…”

O-Linked N -Acetylglucosaminylation of Sp1 Inhibits the Human Immunodeficiency Virus Type 1 Promoter

“…Primers specifically amplifying recombinant OGT were selected: the forward primer recognized specifically the vector-encoded 5Ј untranslated region of the recombinant OGT mRNA (5Ј-TCCAGTGTGGTGGAATTCTG-3Ј); the reverse primer was homologous to a sequence corresponding to the N-terminal region of both endogenous and recombinant OGT (5Ј-TTGCGTCTCAATTGCTTTCA-3Ј). Amplification of GAPDH was performed using the forward primer 5Ј-AG CCACATCGCTCAGAACAC-3Ј and the reverse primer 5Ј-GAGGCATTGCT GATGATCTTG-3Ј as described previously (44).…”

Modified Vaccinia Virus Ankara Triggers Chemotaxis of Monocytes and Early Respiratory Immigration of Leukocytes by Induction of CCL2 Expression

“…Amplification of CCL2/MCP-1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNAs were performed as described previously (21,25) with the help of 26 amplification cycles. Similarly, amplification of the MVA 078R gene (GenBank accession no.…”