AdeptTect Speed HS PCR Master Mix (dye plus) Ⅱ is a 2-fold pre-mixed product for rapid PCR reaction, featuring fast extension speed (15 sec/kb), high amplification efficiency and high annealing efficiency, which is suitable for amplification of long DNA fragments and DNA fragments with complex structure. It is suitable for the amplification of DNA fragments and complex structures. The PCR reaction can be carried out by adding template, primer and water to the solution; at the same time, the product also contains the coloring reagent required for electrophoresis detection, which makes the product appear purple-red color, and the product can be directly used for agarose gel electrophoresis after the PCR reaction is completed; it is easy to operate, which can minimize the human error and obtain the results of the test in a relatively short 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 high elongation speed (up to 15 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.
④ Suitable for amplification of long fragments, 30 kb DNA fragments can be amplified using Human gDNA as template.
⑤ Suitable for amplification of complex DNA fragments such as high GC DNA fragments.
Product Composition
individual parts making up a compound
norm
2X Speed HS PCR Master Mix (dye plus) II
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 to amplify different lengths of DNA fragments, and the results were very good.
The electrophoresis results are shown below:
Example 2
Using gDNA of different species as templates to amplify DNA fragments with different GC contents, high AT and high GC DNA fragments were well amplified.
The electrophoresis results are shown below:
Example 3
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 a template to amplify DNA fragments of different lengths, and very good amplification results were obtained.
The electrophoresis results are shown below:
Example 4
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 4 kb DNA fragments, and the target fragments were amplified with template amounts as low as 1 ng.
The electrophoresis results are shown below:
Example 5
Using Human gDNA as template, this kit can amplify DNA fragments of different lengths, and it can amplify DNA fragments of 30 kb very well.
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.
