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Laser Cryo-Tag® Cryo-Babies® Sheets

Item #: ASLCRYSTS 
Pfu DNA Polymerase

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Pfu DNA Polymerase 
Pfu DNA polymerase is a heat stable DNA polymerase which has 5' 3' DNA polymerase and 3' 5' exonuclease (proofreading) activities. Pfu DNA polymerase retains the high fidelity, sensitivity and processivity with an error rate six-fold lower than Taq DNA polymerase, and significantly lower than the error rates of most other proofreading enzymes or DNA polymerase mixtures (1). This product is supplied with the unique Intact Genomics 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM, and 5X Magic Enhancer that enables efficient amplification of GC rich templates up to 84%. The physical purity of this enzyme is 98% as assessed by SDS-PAGE with Coomassie® blue staining.

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Product Source
E. coli strain expressing a Pfu DNA Polymerase gene from Pyrococcus furiosus.

Contents & Storage

  1. Pfu DNA Polymerase
  2. 10x PCR Buffer with Mg²+
  3. 5x Magic Enhancer
  4. 10 mM dNTP (Cat. # 3312d, 3314d only)

Store all contents at -20 °C.

Storage Buffer 
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C.

10X PCR Buffer with Mg2+ 
100 mM Tris-HCl pH 9.0, 15 mM MgCl, 100 mM KCl, 80 mM (NH4)2SO4, 0.5% Igepal CA 630

Unit Definition 
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTP into acid-insoluble form in 30 minutes at 72 °C.

Protocol
1. Thaw 10x PCR Buffer, dNTP mix, primer solutions, 5x Magic Enhancer (if required) and mix thoroughly before use. 
2. Prepare a reaction mix according to the following table: The reaction mix typically contains all the components needed for PCR except the template DNA.

 

PCR reaction set up:

Template DNA

xµl (0.01-0.5µMg)

10x PCR Buffer

10.0µl

dNTP (10 mM)

2.0µl

Forward Primer

xµl (0.1- 0.5µMM)

Reverse Primer

xµl (0.1- 0.5µMM)

5x Magic Enhancer (optional)

20µl

Pfu DNA Polymerase (5 U/µl)

0.5µl

H2O up to

100.0µl

3. Mix the reaction mixture thoroughly.
4. Add template DNA to the individual PCR tubes containing the reaction mixture. 
5. Program the thermal cycler according to the manufacturerâs instructions. A typical PCR cycling program is outlined in the following table.

PCR cycling conditions:

Steps

Temp.

Time

Cycles 

Initial denaturation

95 °C

3-5 min

1

Denaturation

94 °C

30-60 sec 

  

25-35

Annealing

52-66 °C

30-60 sec 

Extension

72-74 °C

1-2 min 

Final extension

72-74 °C

10 min 

1

Hold 

4-12 °C

â

6. Place the PCR tubes in the thermal cycler and start the cycling program.

PNPCRREAG

Magnesium
Mg
Dye
No Dye
Enzymes
Intact Genomics
Pkg
Indiv
Type
Proofreading
 
Item#:
ASPRIMADNAPOL
 
SKU Number Color Qty Size Temp Usage Type Price Quantity Add to Cart
LCRY-2380 White 2380/pk 0.94 x 0.5 In. -196 - 150 C Cryo-Babies
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LCRY-2380-R Red 2380/pk 0.94 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-2380-O Orange 2380/pk 0.94 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-2380-Y Yellow 2380/pk 0.94 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-2380-G Green 2380/pk 0.94 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-2380-B Blue 2380/pk 0.94 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1400 White 1600/pk 1 x 1 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1700 White 1700/pk 1.28 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1700-R Red 1700/pk 1.28 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1700-O Orange 1700/pk 1.28 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1700-Y Yellow 1700/pk 1.28 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1700-G Green 1700/pk 1.28 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1700-B Blue 1700/pk 1.28 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-2500 White 2,500/pk 2.5 x 0.5 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-3020 White 240/pk 3 x 2 In. -196 - 150 C Cryo-Babies
67.00
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LCRY-1300 White 840/pk 1.125 x 1.75 In. -196 - 150 C Cryo-Tag
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LCRY-1200 White 1,200/pk 1.50 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1200-R Red 1,200/pk 1.50 x 0.75 In. -196 - 150 C Cryo-Tag
60.00
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LCRY-1200-O Orange 1,200/pk 1.50 x 0.75 In. -196 - 150 C Cryo-Tag
60.00
Each
LCRY-1200-Y Yellow 1,200/pk 1.50 x 0.75 In. -196 - 150 C Cryo-Tag
60.00
Each
LCRY-1200-G Green 1,200/pk 1.50 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
Each
LCRY-1200-B Blue 1,200/pk 1.50 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1100 White 1,040/pk 1.69 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1100-R Red 1,040/pk 1.69 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
Each
LCRY-1100-O Orange 1,040/pk 1.69 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
Each
LCRY-1100-Y Yellow 1,040/pk 1.69 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
Each
LCRY-1100-G Green 1,040/pk 1.69 x 0.75 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1100-B Blue 1,040/pk 1.69 x 0.75 In. -196 - 150 C Cryo-Tag
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LCRY-1258 White 600/pk 2.625 x 1 In. -196 - 150 C Cryo-Tag
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LCRY-1258-R Red 600/pk 2.625 x 1 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1258-O Orange 600/pk 2.625 x 1 In. -196 - 150 C Cryo-Tag
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LCRY-1258-Y Yellow 600/pk 2.625 x 1 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1258-G Green 600/pk 2.625 x 1 In. -196 - 150 C Cryo-Tag
67.00
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LCRY-1258-B Blue 600/pk 2.625 x 1 In. -196 - 150 C Cryo-Tag
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LCRY-1000 White 20/pk 8.5 x 11 In. -196 - 150 C Cryo-Tag
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Laser Cryo-Babies White
Laser Cryo-Tag® Cryo-Babies® Sheets
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