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FlaPure Mini Plasmid Kit (PE707)

FlaPure Mini Plasmid Kit (PE707)

Improved SDS alkali lysis method, high-purity plasmid can be extracted in 30 min.

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Product Description

Improved SDS alkali lysis method, high-purity plasmid can be extracted in 30 min.




Highlights


1) Easy to operate: Plasmid DNA extraction from multiple samples can be completed within 30 min.

2) High efficiency: more than 85% of plasmid DNA can be extracted from bacterial cells.





Introduction  


    This kit adopts the improved SDS-alkali lysis method, combined with the advanced silica gel membrane adsorption technology, to achieve the purpose of rapid purification of plasmid DNA. It is suitable for extracting up to 20 μg of high-purity plasmid DNA from 1~4 mL of bacterial culture. The extracted plasmid DNA can be used for molecular biology experiments such as enzyme digestion, PCR, sequencing, bacterial transformation, in vitro transcription and translation, etc.





Composition


Contents

Amount(200 rxns)

RNase A(100 mg/mL)

60 μL

Buffer BL

60 mL

Buffer P1

60 mL

Buffer P2

60 mL

Buffer P3

80 mL

Buffer W1

2×72 mL

Elution Buffer

25 mL

EC

200




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Q1Why is plasmid DNA yield low?

A1

1)  Plasmid copy number is low. Vectors can cause significant fluctuations in plasmid yield due to differences in copy number. High-copy-number vectors often fluctuate by 2 to 3 times the yield (3~16 μg of high-copy-number plasmid vectors per milliliter of bacterial culture cultured overnight). Long fragment plasmids and expression vectors are often dominated by medium and low copy numbers, and the yield per milliliter of bacterial solution is about 0.5~2 μg.

Low-copy plasmids: pBR322, pACYC and its derivatives, pSC101 and its derivatives, SuperCos, pWE15.

High copy plasmids: pTZ, pUC, pBS, pGM-T.

2) Bacteria problem. Plasmids are lost in the process of strain preservation. It is best to streak and activate before culturing bacteria to stabilize the yield.

3) Bacteria are not sufficiently lysed. Bacteria must be fully resuspended in Buffer P1/RNase A, as agglomerated bacteria cannot be lysed, which will reduce the yield..

4) The reagent preparation is wrong. If there is precipitation in Buffer P2, it needs to be heated to dissolve, and the volume of ethanol added to Buffer W1 is not accurate.

5) The elution efficiency is low. Preheat the Elution Buffer to 60~65℃, and perform the second elution.

 

Q2Why is there genomic DNA contamination in plasmid DNA?

A2

1) 1) The bacterial culture time is too long. Bacterial liquid culture time should be controlled within 12~16 h.

2) 2) The cracking problem. When adding Buffer P2, it must be mixed by gentle inversion; when processing multiple samples, the total time should not exceed 5 min after adding Buffer P2.



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