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Nexus RT-PCR Premix prealiquoted (50ulx96)
 
  Quantity in Basket: None
Code: BNRTP501
Price: $485.00
Shipping Weight: 0.50 pounds
 
 
Quantity:
 
Untitled Document
NexusRT-PCR Premix  
DESCRIPTION
Bionexus RT-PCR Premix is optimized ready-to-use mixture of M-MLV Reverse Transcriptase RNase H Minus, ExActTM Taq, reaction buffer, dNTP mixture, stabilizer and tracking dye as 2 fold concentration.
It can be provided as 5X mastermix pre-aliquoted into each PCR tube, thus PCR reaction simply can be preformed by only adding primers and template DNA reducing any mistake and contaminations, and making a high reproducibility with a various and large number of samples.
 
CONTENTS

Contents
Reaction Vol. Final Conc.
20 ul
50 ul
M-MLV Reverse Transcriptase
120 U
300 U
ExActTM Taq
1.5 U
3.75 U
Each dNTP (dATP, dCTP, dGTP, dTTP)
400 µM
400 µM
MgCl2
1.5 mM
1.5 mM
Stabilizer & tracking dye
-
-
GENERAL PROTOCOL
1. Heat the template RNA and primer at 65Cº for 5 min and quick chill on ice (or perform after step 3)
2. Add the heated template RNA and primer
3. Add DEPC-treated sterile water up to 20 ul or 50 ul
4. Incubate at 42Cº (up to 55Cº) for 1 hr
5. Perform PCR
5.1. Heat denaturation at 96Cº for 3 min
5.2. PCR cycle (25~35 cycles)
94Cº 1 min
55Cº1 min
72Cº 1 min
5.3. Last extension at 72Cº for 5 min
6. Load PCR reaction samples on agarose gel and perform electrophoresis

Concentration of template RNA and primer

     20 ul reaction

50 ul reaction
Total RNA
1 ng - 5 ug
1 ng - 5 ug
mRNA
1 ng - 0.5 ug
1 ng - 0.5 ug
Specific primer
0.5 -0.1 uM
0.5 - 1 uM
 
STORAGE CONDITION
-20 Cº
 

Fig.1. Amplification of various cDNAs using Bionexus's RT/PCR Premix with human A49 cell line total RNA. M: 1 kb ladder, Lane 1 : TGF-beta 2 (1258bp), Lane 2 : TGF-beta 3 (1262bp), Lane 3 : Phospholipase 2A (449 bp).
Fig.2. cDNA (HDPE) Amplification of HeLa cells using RT/PCR Premix at different cDNA synthesis temperature. M: 1 kb ladder, Lane 1 : 45 Cº, Lane 2 : 50 Cº
Fig.3. cDNA (TNFR) Amplification of HeLa cells using RT/PCR Premix with serially diluted total RNA from 1ug to 0.1ug M: Lambda Hind III
 
Products
Cat#
Size
Nexus RT-PCR Premix, 2x mastermix
BNRTP700
1ml
Nexus RT-PCR Premix, 5x mastermix, prealiquoted, 96 tubes
BNRTP500
20ulx96
Nexus RT-PCR Premix, 5x mastermix, prealiquoted, 96 tubes
BNRTP501
50ulx96


Related Item(s)
Code Name Price  
BNRTP700 Nexus RT- PCR Premix (1ml) $175.00
BNRTP500 Nexus RT-PCR Premix prealiquoted (20ulx96) $285.00