Invitrogen™ Dynabeads™ mRNA DIRECT™ Purification Kit

The Dynabeads™ mRNA DIRECT™ Kit is designed for simple and rapid isolation of pure, intact polyadenylated (polyA) mRNA directly from the crude lysate of animal and plant cells and tissues.

Supplier:  Invitrogen™ 61012

Catalog No. 61-012

Includes: 10mL Dynabeads oligo (dT)₂₅, 60mL Lysis/Binding Buffer, 120mL Washing Buffer A, 60mL Washing Buffer B, 15mL 10mM Tris-HCl (Elution buffer), Contains enough reagents for 40 standard isolations.

Description

The Dynabeads™ mRNA DIRECT™ Kit is designed for simple and rapid isolation of pure, intact polyadenylated (polyA) mRNA directly from the crude lysate of animal and plant cells and tissues. The isolated mRNA is suitable for use in all downstream applications.

Advantages of the Dynabeads mRNA DIRECT Kit:
• Fast—15 minute procedure yields pure, intact mRNA
• Highly pure mRNA isolation—best choice upstream of cDNA synthesis
• Sensitive mRNA isolation—enables cDNA synthesis and cDNA library construction from ultra-small starting samples (enables cDNA library construction from a single cell)

System overview
The isolation protocol relies on base pairing between the polyA residues at the 3' end of most mRNA, and the oligo (dT)₂₅ residues covalently coupled to the surface of the Dynabeads. Other RNA species lacking a polyA tail will not hybridize to the beads and are readily washed away. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded. RNase inhibiting agents in the Lysis/Binding Buffer together with stringent hybridization and washing conditions ensure the isolation of pure, intact mRNA from crude samples rich in RNase, without the use of strong chaotropic agents. The mRNA purification beads specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples (see protocol). 1 mg of Dynabeads Oligo (dT)₂₅ beads (200 μL) binds up to 2 μg of mRNA. A typical mammalian cell contains about 10–30 pg of total RNA, from which 1–5% is mRNA.

Order Info

Shipping Condition: Room Temperature

Specifications

• Content And Storage:

  • 10 mL Dynabeads Oligo (dt)₂₅, 4°C
  • 60 mL Lysis/Binding Buffer; 4°C
  • 120 mL Washing Buffer A; 4°C
  • 60 mL Washing Buffer B; 4°C
  • 15 mL 10 mM Tris-HCl; 4°C
  
  

• Elution Volume: 10 to 100 μL
• Final Product Type: mRNA
• For Use With (Equipment): Automated Liquid Handling Systems
• Green Features: Beads may be reused for multiple extractions
• Shipping Condition: Room Temperature
• Yield: 2 μg mRNA per 200 μL of beads (Binding capacity)

• Isolation Technology: Magnetic Bead
• Sample Type: Cells, Plants, Plasma, Serum, Tissue, Total RNA, Blood
• High-throughput Compatibility: High-throughput Compatible
• For Use With (Application): RT-PCR, qPCR, cDNA library construction, microarray analysis
• Quantity: 10 mL
• Starting Material Amount: Cells: ≤10⁶
  Plant: ≤400 mg
  Tissue: ≤200 mg

For Research Use Only. Not for use in diagnostic procedures.