Low dsRNA

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**T7 RNA Polymerase: Recombinant Protein for In Vitro RNA Synthesis**


**Overview:**

T7 RNA polymerase is an enzyme derived from recombinant protein expression in *Escherichia coli*. It is widely used for in vitro transcription due to its high specificity and efficiency in synthesizing RNA from a DNA template with a T7 promoter sequence. The enzyme catalyzes the 5'→3' synthesis of RNA, utilizing nucleoside triphosphates (NTPs) as substrates.


**Promoter Sequence:**

The T7 promoter sequence required for transcription initiation is:

5'-TAATACGACTCACTATAG*-3'

(*Note: G* indicates the first base of the RNA transcript.*)


**Template Compatibility:**

- DNA templates with double-stranded linear blunt ends or 5' protruding ends.

- Linearized plasmids and PCR products can serve as templates for T7 RNA polymerase.


**GMP Compliance:**

This product is manufactured following Good Manufacturing Practice (GMP) regulations, ensuring high quality and consistency. It is provided in liquid form.


**Features:**

- Capable of synthesizing both long and short RNA transcripts.

- High yield: Produces 100-200 μg of RNA from 1 μg of DNA template.

- Easy to operate with high efficiency.

- Low endotoxin levels.

- Tested to confirm the absence of endonucleases, exonucleases, and RNases.


**Applications:**

- Preparation of radiolabeled RNA probes.

- RNA generation for studies of RNA structure, processing, and catalysis.

- Expression control via antisense RNA.

- Synthesis of mRNA and single-guide RNA (sgRNA).


**Protocol Highlights:**

1. **Template Preparation:**

   - Use linearized plasmids or PCR products with a T7 promoter sequence.

2. **Transcription Reaction Setup:**

   - Mix the DNA template with T7 RNA polymerase, NTPs, and other necessary transcription components.

3. **Incubation:**

   - Allow the reaction to proceed under optimal conditions for RNA synthesis.

4. **RNA Purification:**

   - Purify the synthesized RNA to remove any residual enzymes and unincorporated nucleotides.


By following the detailed protocol, users can efficiently produce high-quality RNA suitable for various research applications.