这是个完整强大的mRNA扩增试剂盒mRNA amplification kit ，优化后可以使用50ng至1微克的总RNA作为起始材料。该试剂盒可以生成5至50微克放大的mRNA用于标记。并可以与总RNA提取试剂盒以及氨基烯丙基标记试剂盒连用。

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.