Notes: A PCR product amplified from a mutant RB (retinoblastoma protein) cDNA was cloned into the pCI-neo Mammalian Expression Vector. In vivo cyclin-mediated phosphorylation was assayed by cotransfecting the RB expression plasmids with members of the cyclin D or cyclin E family into an RB (-/-) cell line. (0571)

Notes: Genomic DNA was purified from blood or cultured fibroblasts, by means of the Wizard^® Genomic DNA Purification Kit. Total RNA was isolated from cultured fibroblasts by use of the SV Total RNA Isolation System, and it was amplified by Access RT-PCR System by use of exonic primers. An expression vector containing an S-tag attached to the 5´ end of the FALDH cDNA was constructed using pCI-neo Mammalian Expression Vector. (0482)

Notes: The PolyATtract^® System 1000 was used to isolate poly A+ RNA directly from bovine retinas. The isolated RNA was used for RT-PCR amplification. (0369)

Notes: The RNAgents^® Total RNA Isolation System was used to isolated total RNA from Porphyromonas gingivalis cultures. The isolated RNA was used for RT-PCR. The Prime-a-Gene^® Labeling System was used to make probes for Southern analysis. (0366)

Notes: Genomic DNA was isolated from Nectria haematococca mycelia with the Wizard^® Genomic DNA Purification Kit using a slightly modified protocol.  Mycelia were ground in liquid nitrogen and lyophilized before lysing in a modified lysis buffer consisting of 50mM Tris/HCl, pH 7.5, 50 mM EDTA (pH 8.0), 3% SDS and 1% mercaptoethanol.  The isolated DNA was the template for PCR. (0158)

Notes: This paper describes a quick method for analyzing plant microsatellites, also referred to as Simple Sequence Repeats (SSRs), using adaptor-ligated genomic DNA, PCR, and the Streptavidin MagneSphere^® Paramagnetic Particles. Briefly, 200μg of Streptavidin MagneSphere^® Paramagnetic Particles (pre-equilibrated with 6X SSC) were added to 100ng of a PCR product annealed to a biotinylated-repeat oligo. The captured fragments were washed, size-fractionated and used in PCR with AP-11 primers. (3050)

Notes: cDNAs for the receptor genes were obtained by RT-PCR using the Access RT-PCR System. (0569)

Notes: The size of PCR products were determined in relation to the PCR Markers. (1061)

Notes: pGEM^®-T Easy Vector Systems (2068)

Notes: A minigene was constructed with five exons and four introns from exons 5-9 of the CYP27 gene. The 2111bp product was generated with primers containing a start codon and a stop codon. The product was directly cloned into the pTARGET™ Mammalian expression vector and transfected into COS-1cells. RNA was isolated from these cells and RT-PCR was performed to amplify the spliced product. This assay could distinguish between normal splicing of the third intron and aberrant splicing of due to the Arg362Ser mutation. The proteins produced by the normal and mutant minigenes were also assessed by immunoblotting. (1333)

Notes: PCR products were purified by Wizard^® PCR Preps DNA Purification System and sequenced by use of the fmol^® DNA Cycle Sequencing Systems. (0247)

Notes: To confirm the G to A mutation caused alternative splicing, a minigene containing exon 6 with or without the G to A mutation was amplified. The 2111bp minigene was T/A cloned into the pTargeT™ Mammalian Expression Vector and transfected into COS cells. The RNA was isolated from the transfected cells and analyzed by RT-PCR. The G to A mutation resulted in a truncated RT-PCR product. To confirm the mutation would cause a truncated, inactive protein. A cDNA was constructed that would be the result of the alternative splicing and subcloned into the pTargeT™ Vector. COS cells expressing the mutant had no detectable sterol 27 hydroxylase activity. (1330)

Notes: The authors used RQ1 RNase-Free DNase to remove contaminating genomic DNA from total RNA samples. Cu, Zn, and MN SOD mRNA sequences were amplified from hippocampal neuronal RNA using the Access RT-PCR System. (2218)

Notes: PCR was used to amplify the VHL coding and promoter region in six fragments from genomic DNA, and SSCP analysis was performed to detect intragenic mutations. Fragments producing an aberrant SSCP band were purified using the Wizard^® PCR Preps DNA Purification System and subsequentially sequenced. (0176)

Notes: Total RNA was isolated from rat aorta and iliac arteries with the RNAgents® Total RNA Isolation System. The isolated RNA was used for RT-PCR. (1379)

Notes: The Access RT-PCR System was used to determine the presence of hepatitis C virus RNA in chimpanzee serum. (1465)

Notes: The pGEM® -T Easy Vector Systems, Wizard® PCR Preps DNA Purification System and Wizard® Plus Minipreps DNA Purification Systems were used in this study. (1959)

Notes: The S100A2 protein cDNA was subcloned into the pGEMEX^®-2 Vector and expressed in BL21(DE3)pLysS E.coli cells. The protein was purified by a referenced method. (1173)

Notes: The Access RT-PCR System was used to compare the abundance of GnT-IV mRNA in various bovine tissues. (0694)

Notes: Luciferase reporter constructs were cotransfected into 3T3-L1 preadipocytes with the pRL-CMV Vector at a 25:1 ratio and luciferase activities were determined with the Dual-Luciferase^® Reporter Assay System. RT-PCR was accomplished with AMV Reverse Transcriptase and Taq DNA Polymerase. RNase Protection Assay were performed with the aid of the RNase ONE™ Ribonuclease. (0988)

Notes: In this paper, Tfl DNA polymerase was used for two-step RT-PCR. The amplified fragments were cloned into the pGEM®-T Vector. Promega's Terminal Deoxynucleotidyl Transferase (TdT) was used to end label specific oligos for screening a frog brain cDNA library. T7 and T3 RNA polymerases were used to make digoxigenin labeled riboprobes for in situ hybridization. (1281)

Notes: The pGEM®-T Easy Vector was used to clone PCR products amplified from the prophage inserts of various lysogenic Lactococcus strains. (0193)

Notes: The Wizard® Genomic DNA Purification Kit was used to purify genomic DNA from Haemophilus influenzae. The isolated DNA was used for amplification. (1310)

Notes: For the RT-PCR experiment, total RNA isolated from fly heads was reverse transcribed by Promega AMV-RT and then the cDNA was amplified by Promega Taq DNA polymerase. (0574)

Notes: Total RNA was reverse-transcribed with random hexamer primers and AMV Reverse Transcriptase and then amplified for sequence analysis by nested PCR. Poly(A)+ RNA was purified with the PolyATtract^® mRNA Isolation System. (0382)