Amplification & PCR Kit

''MiniAmp'' mRNA Amplification Kit

Description

This complete and robust mRNA amplification kit is optimized to use 50ng to 1µg of total cellular RNA as starting material. This kit can generate 5 to 50 µg of amplified mRNA ready for labelling. It can be used in conjunction with the Total RNA extraction kit and with the AminoAllyl labelling kit.

Reference Designation
MAK MiniAmp mRNA Amplification Kit (10 reactions)

Specifications & Protocol

Needed Equipement

• Ethanol 100%
• Ethanol 70%
• Ice for incubation
• PCR tubes
• Thermal cycler
• Refrigerated microfuge capable of 10,000 rpm or more
• Micropipettors
• Vortex mixer
• Homogenizer (if using tissue)
• Temperature regulated water bath or dry bath

Kit Contents

Items Packaging Storage
 Enzyme Mix 1  16.5µl in 0.5ml tube -20°C Freezer
 RT Booster (enhancer solution)  50µl in 0.5ml tube -20°C Freezer
 Reaction buffer 1  50µl in 0.5ml tube -20°C Freezer
 dNTP Mix  40µl in 0.5ml tube -20°C Freezer
 DTT  50µl in 0.5ml tube -20°C Freezer
 Primer 1 Mix  11µl in 0.5ml tube -20°C Freezer
 Primer 2  11µl in 0.5ml tube -20°C Freezer
 Primer 3  22µl in 0.5ml tube -20°C Freezer
 Primer 4  11µl in 0.5ml tube -20°C Freezer
 Enzyme Mix 2  11µl in 0.5ml tube -20°C Freezer
 Reaction Buffer 2  150µl in 0.5ml tube -20°C Freezer
 DNA Binding Buffer  1.1ml in 2ml tube Room Temp
 DNA Wash Buffer Concentrate  0.5ml in 8ml bottle Room Temp
 DNA Micro Column  10 Room Temp
 Elution Tube (1.5ml)  10 Room Temp
 2 ml Wash Tube  10 Room Temp
 Nuclease Free Water  1.5ml in 2ml tube -20°C Freezer
 Enzyme Mix 3  22µl in 0.5ml tube -20°C Freezer
 NTP Mix  79.2µl in 0.5ml tube -20°C Freezer
 Reaction Buffer 3  22µl in 0.5ml tube -20°C Freezer
 DNAse  11µl in 0.5ml tube -20°C Freezer
 Sodium Acetate Solution  50µl in 0.5ml tube -20°C Freezer
 Linear Acrylamide  20µl in 0.5ml tube -20°C Freezer

Short Protocol

1. Spin all tubes 5 to 10 s before use.
2. Perform first strand cDNA synthesis.
3. Perform second strand cDNA synthesis.
4. Amplify the double-stranded (ds) cDNA.
5. Purify the double-stranded cDNA.
6. Transcribe the ds cDNA with T7 RNA polymerase in vitro.
7. Purify the amplified RNA (aRNA).
8. Perform cDNA synthesis on aRNA using aminoallyl-labeled dNTPs.

Please contact us for further information about the complete protocol.