[TP1260] ExcelTaq™ 5X Fluorescent PCR Master Mix, 200 Rxn

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Description

The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.

 

Features

  • 5’→3’ DNA polymerase activity

  • No detectable 3'→5' exonuclease (proofreading) activity

  • Generates PCR products with 3'-dA overhangs

  • High yield PCR 

  • High reproducibility

  • Reduced  pipetting errors

  • For direct loading into electrophoresis analysis

  • DNA bands can be visualized directly under UV or 470 nm blue light illumination


Applications 

  • Routine PCR

  • Colony PCR

  • High throughput PCR

  • Amplification of DNA fragments up to 8 kb

  • Generation of PCR products for TA cloning

  • DNA labeling

Storage

Protected from light and avoid multiple freeze/thaw cycles
4°C  for     6 months
-20°C  for  24 months

Odoo - Sample 1 for three columns

Elongation capability

λDNA was amplified with ExcelTaq™ 5X Fluorescent PCR Master Mix. The λDNA was amplified with specific primers for amplifying different ranges. The pictures were captured under B-BOX™ blue light as seen in the left picture. While using UV light, the amplicons can be seen in the right picture (M: DM 3160).

Odoo - Sample 1 for three columns

Colony PCR

Colony PCR with ExcelTaq™ 5X Fluorescent PCR Master Dye Mix. The JM109 or modified JM109 was used for amplifying the genomic galactosidase Z gene. The pictures were captured under B-BOX™ blue light as seen in the left picture. While using UV light, the amplicons can be seen in the right picture (M: DM 3160).

Contents

Component

Volume

ExcelTaq™ 5X Fluorescent PCR Master Mix

2 x 1 ml


Storage

Protected from light and avoid multiple freeze/thaw cycles
4°C  for     6 months
-20°C  for  24 months

   

Recommended PCR Condition

Template

1 – 150 ng

Forward primer

0.1 – 0.5 µM

Reverse primer

0.1 – 0.5 µM

5X Fluorescence PCR Master Mix

10 µl

H2O

to 50 µl

Total volume

50 µl


Recommended PCR Program

Steps

Temp.

Time

Cycles

Template denature

94°C

2 min

1

Denature

94°C

30 sec

25-40

Annealing

50-68°C*

30 sec

Extension

72°C

30 sec/kb

Final extension

72°C

1 min

1

*Optimal PCR condition varies according to primers’ thermodynamic properties.

Odoo - Sample 3 for three columns

B-BOX™ Blue Light LED Epi-illuminator

  • 470 nm long wavelength

  • Improved cloning efficiency 

  • Compact, light-weight, and portable (less than 1 kg)

  • Adjustable and removable filter plate allows for gel cutting, visualization, and documentation

Odoo - Sample 1 for three columns

FluoroBand™ DNA Ladder series

  • Sharp bands

  • Quick reference enhanced bands 

  • Ready-to-use— premixed with loading dye for direct loading

  • Stable— room temperature storage over 6 months

  • Directly observed by UV or blue light— premixed with high sensitive fluorescent dye

Odoo - Sample 3 for three columns

FluoroVue™ Nucleic Acid Gel Stain

  • Excellent for in-gel staining

  • Sensitivity up to 0.14 ng (DNA) or or 1 ng (total RNA)

  • A safe alternative to EtBr

  • Suitable for blue or UV light

Odoo - Sample 1 for three columns

High Fidelity PCR amplification

Amplification of target gene with HiFi™ DNA polymerase to minimize error rate.

Odoo - Sample 2 for three columns

Gel electrophoresis

Staining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency.   

Safe fluorescent dyes

Blue-light illuminator

Odoo - Sample 3 for three columns

Ligation

Blund-end PCR amplicons can directly ligate with PCR cloning vector.  

Odoo - Sample 3 for three columns

Transformation

Prepare competent cells with high efficiency and transform with time-saving protocol.