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PREMIUM Bullseye Taq DNA Polymerase Master Mix

Item #: ASPCRREAG3 

Suggested Protocol using Taq 2x Mix

This protocol serves as a guideline for primer extensions. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined.


Notes:

Set up reaction mixtures in an area separate from that used for DNA preparation or product analysis. Work on ice at all times.

The final MgCl2 concentration of this 2x Master Mix is 1.5 mM. If more than 1.5 mM MgCl2 is required, use 25 mM MgCl2 (may be purchased separately) to adjust the Mg2+

 

  

Additional volume (µl) of MgCl2 per 50 µl reaction

Final MgCl2 conc. in reaction (mM)

1.5

2

2.5

3

3.5

4

4.5

Additional volume of 25 mM MgCl2 per reaction (µL):

0

1

2

3

4

5

6

 Thaw Taq 2 Master Mix and primers. It is important to thaw the solutions completely and mix thoroughly before use to avoid localized concentrations of salt. Keep all components on ice.

 

 The table below shows the reaction set up for a final volume of 50 m If desired, the reaction size may be scaled down. Use 10 µl of the Taq 2x Master Mix in a final volume of 20 µl.

 

 Important: Spin Taq Master Mix vials briefly before use.

 

  

Set up each reaction as follows:

Component

Vol./reaction

Final Conc.

Taq 2x Master Mix

25 µL

1X

25 mM MgCl2

0 µl (0-7 µl)

1.5 mM (1.5-5 mM)

Primer A

1 µL (0.5-5 µl)

0.2 µM (0.1–1.0 µM)

Primer B

1 µL (0.5-5 µl)

0.2 µM (0.1–1.0 µM)

Distilled Water

Variable

– – – –

Template DNA

Variable

Genomic DNA 50 ng (10-500 ng) Plasmid DNA 0.5 ng (0.1-1 ng) Bacterial DNA 5 ng      (1-10 ng)

TOTAL volume

50 µL

– – – –

 

Mix gently by pipetting the solution up and down a few times.

Program the thermal cycler according to the manufacturer´s instructions. For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.

Place the tubes in the thermal cycler and start the reaction.

  

Three- step PCR Program:

Cycles

Duration of Cycles

Temperature

1

2-5 minutesa

95°C

25-35

20-30 secondsb

20-40 secondsc

30 secondsd

95°C

50-65°C

72°C

1

5 minutese

72°C

 

a  Initial denaturation step. Optional, but recommended for genomic DNA

b Denaturation step: Heating the DNA template disrupts the hydrogen bonds yielding ssDNA.

c Annealing step: Primers anneal to ssDNA template. Typically annealing temperature is 3-5°C below the Tm of the primers.

d Extension/elongation step: Taq DNA polymerase synthesizes a new DNA strand complementary to the DNA template. Extension time depends on length of DNA fragment to be amplified. At optimum temperature the DNA polymerase will polymerize 1000 bases per minute.

 

e Final elongation step: After the last PCR cycle to ensure that any remaining ssDNA is fully extended.

Cryo Labels

Cryogenic Labels

Labeling frozen vials, microtubes, microplates, glass tubes, freezer boxes, plastic bags and other containers can be a challenge when storing in temperatures below 20 degrees Celsius.

We offer with a high-quality adhesive that can stick to already frozen vials and other objects. The adhesive on this type of label will adhere to a surface in wet and/or cold conditions. The adhesive on the label is specifically designed to stick in temperatures below 80 degree Celsius and as low as 196 degrees Celsius. When storing in deep freeze liquid nitrogen or vapor phase liquid nitrogen our Cryo labels will stay in place. MIDSCI's Cryo labels are resistant to smearing without any additional lamination. They also won't become brittle or cracked.

Our cryogenic labels are made of thermoplastic film and coated with a glove-friendly cryogenic adhesive. This special adhesive let the label to remain strong and sticky so that it can be securely attached to stainless steel, aluminum, paint, powder coated metal and iron.

Providing a high quality of the Cryo label is essential for laboratories, researchers, (etc.). It will provide fewer errors, better accuracy, more proficiency, and all of this leads to better results at a better budget.

These come on a roll ready for your use in a lab!

PNLABEL

Temp Usage
-196°C
SKU Number Vol Magnesium Dye Enzymes Pkg Type Price Available Quantity Add to Cart
BE140303 500 runs Mg No Dye Premium Mix Standard
350.49
Each
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BE180303 500 runs Mg Red Dye Premium Mix Standard
297.88
Each
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Bullseye Taq DNA Polymera
PREMIUM Bullseye Taq DNA Polymerase Master Mix
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