体外转录相关技术

Reaction 1 μl 10X Transcription Buffer (Ambion) 1 μl 10X NTPs (4 mM ATP CTP 1 mM GTP UTP) 2 μl 10 mM GpppG cap (Pharmacia) 2 μl a-UTP (NEN) 800 Ci/mmol 0.2 μl RNasin (Promega) 1 μl ...

Reagents/Solutions DNA (PCR or plasmid) with bacteriophage RNA polymerase promoter in suitable orientation (see note 1) ~1μg/μl in TE buffer 10mM MgCl2 5x Transcription Buffer 0.2M Tris - HCl p ...

Overview With this protocol transcripts that were initiated from specific genes by RNA polymerases prior to permeabilization can be measured. Instead of a nuclear extract permeabilized cells are ...

High-throughput protein analysis integrating bioinformatics and experimental assays. Nucleic Acids Res. 2004 Feb 3;32(2):742-8 The wealth of transcript information that has been made publicly availa ...

Overview With this protocol transcripts that were initiated from specific genes by RNA polymerases prior to permeabilization can be measured. Instead of a nuclear extract permeabi ...

In vitro transcription with yeast nuclear extract Steve Hahn Wear gloves throughout use RNAse free solutions (either autoclaved or sterile filtered) and clean bench and pipetmen with 95% ethanol befo ...

Polymerase III in vitro Transcription Steve Hahn last modified 10/15/99 For the following reactions use appropriate shielding and dispose of radioactive waste properly! A 20 microliter transcription r ...

Steve Hahn Last Modified Fri Apr 25 2003 Wear gloves throughout use RNAse free solutions (either autoclaved or sterile filtered) and clean bench and pipetmen with 95% ethanol before use to eliminate ...

Run-on transcription monitors the regulation of transcription in isolated nuclei. A. Preparation of Nuclei - (do everything at 0℃ to 4℃ in 50 ml tubes) 1. Pellet between 30 and 300 million cells at 15 ...

N.B: Gloves should be worn at all times during preparation of in vitro RNA. All solutions should be RNase-free i.e. made with DEPC water if home-made or bought specifically to use for RNA work. You w ...

1 ml linearized DNA template pBS-SK- ERD (X-E) (0.1-0.5 mg) 7.8 ml DDW 5 ml 5X transcription buffer ( stratagene ) 1 ml 750 mM DTT 1 ml Rnasin (RNase inhibitor; 15U/ml) 1 ml 10 mM ATP (pH 7) 1 ml 10 m ...

Polymerase III in vitro Transcription Steve Hahn For the following reactions use appropriate shielding and dispose of radioactive waste properly! A 20 microliter transcription reaction contains: 4.0 ...

In vitro transcription with yeast nuclear extract Steve Hahn Last Modified Fri Apr 25 2003 Wear gloves throughout use RNAse free solutions (either autoclaved or sterile filtered) and clean bench and ...

The following protocol is for MEGAscript II Kit (Ambion). 1. PCR amplify the DNA template. The 5'-end of the template should contain the minimum promoter sequenences of T7 or Sp6 or T3. A 5-primer wi ...

Transcription of cRNA probe Reagents/Solutions DNA (PCR or plasmid) with bacteriophage RNA polymerase promoter in suitable orientation (see note 1 ) ~1µg/µl in TE buffer 10mM MgCl2 5x Tr ...

in vitro Transcription Reaction 1 µl 10X Transcription Buffer (Ambion) 1 µl 10X NTPs (4 mM ATP CTP 1 mM GTP UTP) 2 µl 10 mM GpppG cap (Pharmacia) 2 µl a-UTP (NEN) 800 Ci/mmo ...