Invitrogen™ SuperScript™ VILO™ cDNA Synthesis Kit

Product Code.	No. of Reactions	unitSize
10499763	50 Reactions	50 reactions
10704274	250 Reactions	250 reactions

Product Code. 10499763 Supplier Invitrogen™ Supplier No. 11754050

Description

Includes

5X VILO Reaction Mix (200μL) and a 10X SuperScript III Enzyme Blend (100μL), which contains: SuperScript III RT, RNaseOUT Recombinant Ribonuclease Inhibitor, and a proprietary helper protein

The SuperScript VILO cDNA Synthesis Kit is designed to generate first strand cDNA for two-step RT-qPCR applications. The kit is supplied in a two-tube format with the VILO reaction mix and an enzyme blend in separate tubes.

The SuperScript VILO cDNA Synthesis Kit is designed to generate first strand cDNA for two-step RT-qPCR applications. The kit is supplied in a two-tube format with the VILO reaction mix and an enzyme blend in separate tubes. The enzyme blend contains SuperScript III Reverse Transcriptase (RT), a genetically engineered MMLV RT that has reduced RNase H activity and improved thermostability for highly efficient cDNA synthesis.

The SuperScript VILO cDNA Synthesis Kit can be used to synthesize cDNA from for a wide range of input RNA amounts. This kit provides the high temperature capability of SuperScript III Reverse Transcriptase in an optimized format for generating first-strand cDNA for use in RT–qPCR.

Notes
• For superior first-strand cDNA synthesis in two-step RT-qPCR, try SuperScript IV VILO Master Mix (Cat. No. 11756050) or SuperScript IV VILO Master Mix with ezDNase Enzyme (Cat. No. 11766050).
• This kit is included with EXPRESS Two-Step RT–qPCR kits. Reagents are provided for 50 or 250 20-μL cDNA synthesis reactions.

Order Info

Shipping Condition: Dry ice

Specifications

• Content And Storage:

  • 5X VILO Reaction Mix, 200 μL
  • 10X SuperScript III Enzyme Blend, 100 μL
  
  Store at –5 to –30°C.
  
  

• Detection Method: qPCR
• Format: Kit
• GC-Rich PCR Performance: High
• Reaction Speed: 60 min.
• Technique: Reverse Transcription
• Optimal Reaction Temperature: 42°C
• Quantity: 50 reactions
• Reverse Transcriptase: SuperScript III
• Ribonuclease H Activity: Reduced
• Shipping Condition: Dry Ice
• For Use With (Application): Real Time PCR (qPCR)
• Final Product Type: First-Strand cDNA
• No. of Reactions: 50 Reactions
• Reaction Format: Master Mix
• Reagent Type: Reverse Transcription
• Size (Final Product): Up to 10 kb
• Starting Material: RNA

Frequently Asked Questions (FAQs)

How long can I store the cDNA from my reverse transcription step?

You can store your cDNA at 2-6 degrees C for up to 24 hours. For long-term storage, store the cDNA at -15 to -25 degrees C and add EDTA to a final concentration of 1 mM to prevent degradation.

What is the difference between SuperScript III RT and the RT in the SuperScript VILO kit?

The SuperScript VILO cDNA Synthesis Kit contains a mix of SuperScript III RT and helper proteins which help to increase the efficiency of the reverse transcription reaction and thus improve yield. The RT in the SuperScript VILO kit is active at 42 degrees C due to the helper proteins.

How much of the first-strand cDNA reaction should I load for PCR?

While the volume is dependent on the starting amount of RNA used for the first-strand synthesis and the abundance of the target gene, we'd recommend starting with 10% of the first-strand reaction for your PCR reaction.

In what concentration range of total RNA does the SuperScript VILO cDNA Synthesis Kit (Cat. No. 11754050) work?

Lower limit of RNA concentration: 1.0 pg

Upper limit of RNA concentration: 2.5 µg

What is the RNA input for the Oncomine Focus Assay, Chef-Ready Library kit?

Each reverse transcription reaction requires 10 ng of DNase-treated total RNA (≥1.43 ng/µL).

With the Oncomine Focus Assay, Chef-Ready Library, can I perform reverse transcription using the SuperScript VILO cDNA Synthesis Kit on the Ion Chef System?

Reverse transcription needs to be performed manually in the IonCode 96-well PCR plate, using the SuperScript VILO cDNA Synthesis Kit. SuperScript reverse transcriptase enzyme is not supplied in the kit and must be purchased separately. If the RNA was prepared from FFPE tissue and not previously heat-treated, we recommend heating at 80 degrees C for 10 min, then cooling to room temperature.

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