superscript ii rnase h-rt kit  (Thermo Fisher)

Journal:

Article Title: fbpABC Gene Cluster in Neisseria meningitidis Is Transcribed as an Operon

doi:

Figure Lengend Snippet: (A) The meningococcal fbpABC gene cluster encoding the neisserial iron ABC transporter. The orientations and approximate positions of the gene-specific primers for generating the cDNA and the oligonucleotide primers used in the PCR amplification step of the RT-PCR assays are displayed below the schematic. gDNA, genomic DNA. (B) RT-PCR amplification of total RNA extracted from N. meningitidis B16B6. The PCR amplification products were fractionated in a 1% agarose gel that was stained with ethidium bromide. The ingredients used to generate the amplicon in each lane are depicted in the table above the agarose gel. RT primers are designated as follows: tbp, 48; Asd, 3′HKasdstop; B, 3′fbpBint; and C, 3′fbpCstop. PCR primer pairs are designated as follows: tbpA, 385 and 48; asd, 5′HKasdint and 3′HKasdstop; AB, 5′fbpAint-3′fbpBint; and BC, 5′fbpBint-3′fbpCint. The figure was imaged with a Hewlett-Packard ScanJet HP, edited by using Adobe Photoshop 3.0, and labelled by using Microsoft PowerPoint 97. The sizes of standards in kilobases (lane a) are shown on the left.

Article Snippet: Reverse transcription was performed with the SuperScript II RNase H − RT-PCR kit (Gibco BRL) following the manufacturer's instructions.

Techniques: Amplification, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining