Author: Cobucci-Ponzano, Beatrice; Conte, Fiorella; Benelli, Dario; Londei, Paola; Flagiello, Angela; Monti, Maria; Pucci, Piero; Rossi, Mosè; Moracci, Marco
Title: The gene of an archaeal a-l-fucosidase is expressed by translational frameshifting Document date: 2006_8_18
ID: 69gftii4_23
Snippet: The mRNAs encoding wild-type fucA1 and its various mutants were obtained by in vitro run-off transcription. About 2 mg of each plasmid was linearized with BssHII and incubated with 50 U of T7 RNA polymerase for 1 h 30 min at 37 C. The transcription mixtures were then treated with 10 U of DNAseI (RNAse free) for 30 min. The transcribed RNAs were recovered by extracting the samples twice with phenol (pH 4.7) and once with phenol/chloroform 1:1 foll.....
Document: The mRNAs encoding wild-type fucA1 and its various mutants were obtained by in vitro run-off transcription. About 2 mg of each plasmid was linearized with BssHII and incubated with 50 U of T7 RNA polymerase for 1 h 30 min at 37 C. The transcription mixtures were then treated with 10 U of DNAseI (RNAse free) for 30 min. The transcribed RNAs were recovered by extracting the samples twice with phenol (pH 4.7) and once with phenol/chloroform 1:1 followed by precipitation with ethanol. The mRNAs were resuspended in DEPC-treated H 2 O at the approximate concentration of 0.6 pmol/ml.
Search related documents:
Co phrase search for related documents- ethanol precipitation and phenol chloroform: 1, 2, 3, 4, 5, 6, 7, 8
- phenol chloroform and run transcription: 1
- phenol chloroform and vitro run transcription: 1
- phenol chloroform and wild type: 1
- run transcription and vitro run transcription: 1, 2, 3, 4, 5
- run transcription and wild type: 1, 2
Co phrase search for related documents, hyperlinks ordered by date