AdeptTect Speed HS PCR Master Mix (dye plus) I is a 2-fold pre-mixed product for rapid PCR reaction, with an extension speed of up to 10 sec/kb; it has strong amplification performance and is suitable for amplification of complex templates, and can amplify DNA fragments of up to 10 kb in length; to use it, it is only necessary to add templates, primers, and water to the solution to carry out the PCR reaction. At the same time, the product also adds the coloring reagent required for electrophoresis detection, the reagent is purple-red, and can be used for agarose gel electrophoresis directly after the PCR reaction; the operation is convenient, which can minimize the human error and obtain the test results in a shorter period of time. In addition, monoclonal antibody that can inhibit the activity of DNA Polymerase at room temperature is added to the product, which can be used for Hot Start PCR and effectively inhibit the formation of primer dimer and non-specific amplification.
The 3’ end of the amplified product does not contain the A base, so it cannot be used directly for TA cloning.
Product Advantages
① The product has a fast extension speed (up to 10 sec/kb), which enables the test results to be obtained in a short time.
② This product is a 2X premix, add template, primers and water to carry out the PCR reaction, easy to operate, can minimize human error, greatly improving the efficiency of the experiment.
③ The product contains violet dye, after PCR can be directly carried out agarose gel electrophoresis, without adding electrophoresis sample buffer, electrophoresis has a light violet-red indicator band.
Product Composition
individual parts making up a compound
norm
2X Speed HS PCR Master Mix (dye plus)I
500 μl x 6 pc
Preservation and transportation
Storage temperature: -20℃
Transportation temperature: dry ice or -20℃ ice bag
Example 1
The gDNA of different species was used as templates, and different lengths of DNA fragments were amplified with this kit at an extension rate of 10 sec/kb, and good amplification results were obtained.
The electrophoresis results are shown below:
Example 2
pick E. coli Single colonies were added to 10 μl of RNase free water, mixed well, and then 2 μl of the bacterial solution was used as the template to amplify DNA fragments of different lengths, and all of them were amplified very well.
The electrophoresis results are shown below:
Example 3
Using Human gDNA as a template, different template amounts (500 ng, 200 ng, 100 ng, 10 ng, 1 ng, 100 pg) were added, and the kit was used to amplify 2 kb DNA fragments, and the target fragments were amplified with template amounts as low as 1 ng.
The electrophoresis results are shown below:
table of examples of experiments
Template Type
Amplification speed
Fragment size (kb)
Sequence GC content
Template Extraction Method
template volume
Can it be amplified
Chicken gDNA
10 sec / kb
0.5 kb
41.0%
Magnetic bead method
74 ng
√
Chicken gDNA
10 sec / kb
1.2 kb
36.6%
Magnetic bead method
74 ng
√
Chicken gDNA
15 sec / kb
1.7 kb
68.6%
Magnetic bead method
74 ng
√
Chicken gDNA
10 sec / kb
2.4 kb
49.4%
Magnetic bead method
74 ng
√
Human gDNA
10 sec / kb
1 kb
36.1%
reagent method
100 ng
√
Human gDNA
10 sec / kb
2 kb
38.1%
reagent method
100 ng
√
Human gDNA
10 sec / kb
3 kb
38.7%
reagent method
100 ng
√
Human gDNA
10 sec / kb
4 kb
36.0%
reagent method
100 ng
√
Human gDNA
10 sec / kb
6 kb
40.0%
reagent method
100 ng
√
Human gDNA
15 sec / kb
7.5 kb
37.1%
reagent method
100 ng
√
Human gDNA
15 sec / kb
10 kb
37.7%
reagent method
100 ng
√
Human gDNA
15 sec / kb
12 kb
40.0%
reagent method
100 ng
√
Mouse gDNA
10 sec / kb
1 kb
52.2%
Magnetic bead method
100 ng
√
Mouse gDNA
10 sec / kb
2.6 kb
65.2%
Magnetic bead method
100 ng
√
Mouse gDNA
10 sec / kb
3.4 kb
53.4%
Magnetic bead method
100 ng
√
Mouse gDNA
10 sec / kb
4 kb
49.8%
Magnetic bead method
100 ng
√
Mouse gDNA
15 sec / kb
5 kb
49.8%
Magnetic bead method
100 ng
√
Pig gDNA
10 sec / kb
0.6 kb
47.6%
Magnetic bead method
30 ng
√
Pig gDNA
10 sec / kb
0.8 kb
50.2%
Magnetic bead method
30 ng
√
Pig gDNA
10 sec / kb
2 kb
57.9%
Magnetic bead method
30 ng
√
Pig gDNA
10 sec / kb
2.9 kb
37.7%
Magnetic bead method
30 ng
√
Pig gDNA
15 sec / kb
6 kb
59.3%
Magnetic bead method
30 ng
√
Potato gDNA
10 sec / kb
1.3 kb
39.5%
columnar method
50 ng
√
Potato gDNA
10 sec / kb
2 kb
39.6%
columnar method
50 ng
√
Potato gDNA
10 sec / kb
2.9 kb
38.2%
columnar method
50 ng
√
Potato gDNA
10 sec / kb
5.8 kb
35.4%
columnar method
50 ng
√
Potato gDNA
15 sec / kb
10 kb
38.0%
columnar method
50 ng
√
Rice gDNA
10 sec / kb
1 kb
45.6%
Magnetic bead/column method
100 ng / 200 ng
√
Rice gDNA
10 sec / kb
3 kb
41.5%
Magnetic bead/column method
100 ng / 200 ng
√
Rice gDNA
10 sec / kb
5 kb
40.5%
Magnetic bead/column method
100 ng / 200 ng
√
Rice gDNA
15 sec / kb
7.5 kb
46.8%
Magnetic bead/column method
100 ng / 200 ng
√
Rice gDNA
15 sec / kb
12 kb
44.6%
Magnetic bead/column method
100 ng / 200 ng
√
Tomato gDNA
10 sec / kb
0.5 kb
28.2%
Magnetic bead/column method
120 ng
√
Tomato gDNA
10 sec / kb
1.6 kb
32.2%
Magnetic bead/column method
120 ng
√
Tomato gDNA
10 sec / kb
1.9 kb
36.0%
Magnetic bead/column method
120 ng
√
Tomato gDNA
10 sec / kb
3 kb
37.4%
Magnetic bead/column method
120 ng
√
Tomato gDNA
10 sec / kb
4.4 kb
37.6%
Magnetic bead/column method
120 ng
√
Tomato gDNA
15 sec / kb
5 kb
38.2%
Magnetic bead/column method
120 ng
√
λDNA
20 sec / kb
30 kb
51.1%
–
2 ng
√
Escherichia coli colony
10 sec / kb
1 kb
53.8%
/
single colony
√
Escherichia coli colony
10 sec / kb
2 kb
51.1%
/
single colony
√
Escherichia coli colony
10 sec / kb
3.5 kb
52.2%
/
single colony
√
Escherichia coli colony
10 sec / kb
5 kb
53.2%
/
single colony
√
Escherichia coli colony
10 sec / kb
6.7 kb
51.8%
/
single colony
√
Escherichia coli colony
15 sec / kb
11 kb
50.7%
/
single colony
√
Note: √ indicates able to amplify, / indicates not extracted. -indicates direct purchase.
