We have developed a procedure for preparing extracts from nuclei of human tissue culture cells that directs accurate transcription initiation in vitro from class II promoters. Conditions of extraction and assay have been optimized for maximum activity using the major late promoter of adenovirus 2

the sigma 54-dependent activator FhlA. byspecificity of in vitro transcription activation The nucleotide concentration determines the

We have investigated the effect of in vitro transcription on cruciform stability. Replicative form DNA of 4X1 74 strain ins6240, containing a 48 bp synthetic palindrome in the J-F intercistronic region, was supercoiled in vitro to mean negative superhelical densities (ar) ranging from 0 to 0

Modulating the potency of an activator in a yeast in vitro transcription system.

by transduction of four defined transcription factors. Here, we demonstrate the generation of iPS cells from adult human dermal fibroblasts with the same four factors: Oct3/4, Sox2, Klf4, and c-Myc. Human iPS cells were similar to human embryonic stem (ES) cells in morphology, proliferation, surface antigens

We have developed a simple method to measure RNA synthesis in real time. In this technique, transcription reactions are performed in the pres-ence of molecular beacons that possess a 2¢-O-methylribonucleotide backbone. These probes become ¯uorescent as they hybridize to nascent RNA during

components allow for in vitro implementation of arbitrary circuits. We use only two enzymes in addition to DNA and RNA molecules: RNA polymerase (RNAP) and ribonuclease (RNase). We develop a rate equation for in vitro transcriptional networks, and derive a correspondence with general neural network rate

To elucidate the mechanism of transcription and replication of Sendai virus, we developed an efficient and faithful in vitro transcription system using purified virus particles. The in vitro RNA synthesis was almost entirely dependent on the addition of eukaryotic cell extracts, including those

1. Prepare plasmid DNA containing the protein-coding sequences downstream of a bacteriophage promoter by CsCl or Qiagen or PEG-NaCl methods as the quality of the DNA is crucial for the in vitro transcription step. 2. Digest 10 µg of plasmid DNA with a restriction enzyme that cuts downstream

The in vitro transcription of chick liver chromatin before and after estrogen treatment was studied. Transcription was by endogenous as well as homologous exogenous RNA polymerase II and the products were analyzed for size and specific vitellogenin sequences. Quantitatively more RNA was synthesized