Single Column
InquireCatalog Number｜Packaging
Mat. No |
Ref. No |
No. of preps |
|
4992437 |
GDP116 | 10 |
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Features
● High yield: Up to 1.5 mg of plasmid DNA can be extracted by the direct precipitation of isopropanol.
● High purity: The combination of Buffer P4 and Filtration CS1 can ensure the extraction of high-purity plasmid DNA which can meet the requirements of a variety of transfection experiments.
● Simple and fast: The experiment can be completed in around 1 h with just a few simple centrifugation steps.
● Wide range of applications: The purified plasmids can be applied in molecular biology experiments such as restriction endonuclease digestion, PCR, transformation, sequencing and transfection. The color indicator provided in this kit can ensure the plasmid extraction efficiency.
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Description
The HighPure Maxi Plasmid Kit adopts a unique buffer system incorporation with TIANGEN’s special color indicator to ensure the extraction of large amount of high-purity plasmids. The initial volume of the bacteria sample can be adjusted flexibly according to different experimental requirements. The traditional sopropanol precipitation method is applied to quickly extract large amount of high-purity plasmid DNA, which can be applied in various downstream experiments.
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Kit Contents
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Yield For Reference
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Storage Condition
Room temperature (15-30℃).
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Important Notes
● Add the provided RNase A solution to Buffer P1 (use 1 vial RNase A per bottle Buffer P1), mix, and store at 2-8°C.
● Check Buffer P2 and P4 before use for salt precipitation. If necessary, dissolve the buffer by warming at 37°C for several minutes.
● Prepare about 60 ml of 5 M NaCl buffer.
● Avoid direct contact of Buffer P2 and P4, immediately close the lid after use.
● Draw out the plunger from the Filtration CS1 slowly to avoid membrane loose.
● The amount of extracted plasmid is related to cells concentration and plasmid copy. If working with low copy vectors or large plasmid (>10 kb), it may be beneficial to increase culture volume and to increase Buffer P1, P2, and P4 in proportion. Warm the Buffer TB at 65-70°C before use.
● TIANRed user guide: TIANRed is an indicator, which is harmless and used to make sure that the whole experimental process works well. TIANRed is optional, researchers can determine whether to use according to their experience and experiment purpose. If the purified plasmid is to be used in transfection experiment, TIANRed is not recommended. TIANRed should be mixed with Buffer P1 in the ratio of 1:200 and the color of the mixed solution should be clear red. Add the mixed solution to bacterial cells and the solution would turn turbid red. After that, add Buffer P2 to the turbid solution, the solution would turn clear purple which means a complete lysis. Add Buffer P4 to the purple solution and it would turn clear yellow, which indicate that the neutralization reaction has been done.
Experimental Example
pH Indicator Dye
Figure 1: Buffer P1 containing TIANRed indicator was added to the bacteria culture medium. After thorough mixing, the solution turns into cloudy red.
Figure 2: After adding Buffer P2 and mixing thoroughly, the solution turns into transparent purple. If the cloudy red is present in the purple solution, it indicates that the lysis is insufficient, which will greatly affect the extraction efficiency of the plasmid. Keep mixing the solution until it turns into transparent purple.
Figure 3-4: After mixing Buffer P4 thoroughly, the solution turns into transparent yellow. If purple mix is present in the yellow solution (as shown in Figure 3), it indicates that the renaturation is insufficient, which will result in a decrease in the yield of the double helix plasmid. Keep mixing until the color becomes transparent yellow (Figure 4).
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