CN205133629U - A purifying column for body fluid trace nucleic acid extraction - Google Patents

Abstract

The utility model provides an improve that a purifying column for body fluid trace nucleic acid extraction belongs to the small nucleic acid extraction field of body fluid trace, especially relates to an extraction element structure that is arranged in the minimal small nucleic acid of body fluid content. The utility model provides a can improve a purifying column for body fluid trace nucleic acid extraction of RNA purity. A purifying column for body fluid trace nucleic acid extraction, includes the shell, is equipped with the filter layer in shell inboard lower part, is equipped with cyclic annular clamping ring, its characterized in that in the filter layer top: the outer upper end and interior along constituting outer high interior low inclined plane between the lower extreme of following of cyclic annular clamping ring.

Description

A kind of purification column extracted for body fluid trace dna

Technical field

The utility model belongs to the small nucleic acid extraction field of body fluid trace, especially relates to a kind of improvement of extraction element structure of the small nucleic acid atomic for content in body fluid.

Background technology

RNA is one of most important biological information molecule, an important component part of molecular biology research, no matter be the structure of cDNA library, RT-PCR, RNA sequential analysis, Northern engram analysis, protein In Vitro Translation etc. all depend on high purity global RNA.Therefore, how obtaining high-quality RNA is the problem that researchist pays close attention to.

The universal method that RNA extracts is guanidinium isothiocyanate/phynol method, and its principle is that cell is cleaved under the effect of denaturing agent guanidinium isothiocyanate, the protein denaturation simultaneously on ribosome, and nucleic acid discharges; DNA and RNA discharged due under specific pH condition solubleness difference and lay respectively at mesophase spherule and the aqueous phase of whole system, thus to be separated; Organic solvent extracting, precipitation, obtains purified dna and RNA.Therefore, the RNA rapid extraction reagent be made up of above-mentioned composition forms the basis obtaining high purity global RNA.

At present, the RNA based on above principle market emerging shiploads of merchandise extracts test kit.For purification column, generally comprise lysate, rinsing liquid, wash post liquid, remove RNA enzyme water, remove RNA enzyme adsorption column and remove RNA enzyme collection tube.Efficiently, single-minded in conjunction with the centrifugal adsorbing column of RNA and the buffering system of uniqueness, can from the micro-example of number of different types rapid extraction total serum IgE.

RNA rapid extracting method simplifies the schedule of operation of traditional single stage method, eliminate the step that sex change liquid redissolves and Virahol precipitates again, this not only makes operation simplify, and make RNA lose minimizing, the rate of recovery increases, and make the extracting cycle of RNA foreshorten to 1h by original 4h, this not only saves the time simultaneously, improve working efficiency, and decrease the chance of RNA degraded.And the washing of pillar has the effect of " water displacement wash ", the RNA in pillar does not form agglomerate; In the removal of small molecules thing (salt etc.), liquid residual in centrifugal rear pillar is few.

Although RNA purification column has above plurality of advantages, also there is certain deficiency.After extracting Microrna by above-mentioned means exactly, use conventional nucleic acid and Protein Detection equipment (such as Nanodrop) etc. even to can't detect classical nucleic acid absorbancy peak value (absorption peak at 260nm place), detection obtains value and is generally only <10ng/ul.Its reason cannot be explained always by each major company and research institution, is still changing different chemical reagent, is increasing in original samples amount and carry out studying, improving, to expect to solve the problem; But poor effect.

We finds in experiment, research process, sound is to the basic reason of the purity of RNA: in conventional nucleic acid purification column, the section of gland fixed nucleic acid adsorption film is the structure defect of foursquare ring-type trim ring (as shown in Figure 1, Figure 2), and almost always causing has the elution reagent in partial nucleic acid leaching process residual on trim ring upper surface.Residual elutriant can cause the overall absorbancy lifting of sample in 230-280nm wavelength orientation, masks the concentration of originally very low with regard to detected value nucleic acid, even only can see the photometric absorbance curve of elutriant, and the absorption peak of nucleic acid is masked completely.More seriously, residual elution reagent contains various chemical substance, and when carrying out follow-up polymerize chain reaction amplified reaction (PCR), these reagent can have influence on the efficiency of PCR reaction, even lead to the failure.Extract comparatively in recent years general at microRNA, this elutriant remains the failure of an experiment caused and is still affecting worldwide numerous investigators, make most investigator can only then remove to study cell, the tissue of microRNA rich content.Although increasing company generates to develop successfully can carry out the test kit that in body fluid, miRNA detects, but in actual use, still there is recall rate low, the phenomenon that the abundance of the miRNA obtained that increases is not high enough.Although use pipettor carefully to remove residual elutriant by before in the end Nucleic Acid Elution, the kind recall rate of trace dna in serum can be made to double, and content (copy number) even can increase by 5 powers of 2, about more than 30 times.But this measure causes experimental procedure loaded down with trivial details, waste consumptive material.

But so far, each major company and research institution still do not notice and to extract for micro-microRNA, and the reason causing experimental result bad is just this ring-type trim ring, the extraction column that current domestic and international all companies produce still is using original structure always.

Summary of the invention

The utility model is exactly for the problems referred to above, provides a kind of a kind of purification column extracted for body fluid trace dna improving RNA purity.

For realizing above-mentioned purpose of the present utility model, the utility model adopts following technical scheme, a kind of purification column extracted for body fluid trace dna, comprise shell, side lower part is provided with filtering layer in the enclosure, above filtering layer, be provided with ring-type trim ring, it is characterized in that: the upper end, outer of described ring-type trim ring and interiorly form high outside and low outside scarp between lower end.

As a kind of preferred version, the upper end, outer of described ring-type trim ring and interiorly form tilting plane between lower end.

As another kind of preferred version, the upper end, outer of described ring-type trim ring and the interior cambered surface along forming between lower end to lower recess.

As another preferred version, be provided with liquid discharge pipe in shell lower end; The liquid such as elution reagent are discharged from liquid discharge pipe.

As another preferred version, described filtering layer is sieve plate or filter paper.

As another preferred version, described shell is column plastic casing.

The beneficial effects of the utility model.

The utility model by by section be rectangle ring-type trim ring, be improved to upper end, outer and the interior mode along forming the trim ring on scarp between lower end, the phenomenon that in thorough solution centrifugal process, elutriant is residual, greatly benefit scientific research field, solve researchist and thirst for solving always but fail the technical barrier that succeeds all the time.In addition, the utility model structure is simple, but owing to being in very professional field, very difficultly finds the reason causing and extract residual elutriant in RNA process.Therefore, the utility model extracts the output of product and scientific research result to whole world trace dna, and economic benefit and realistic meaning all can not be despised.

Accompanying drawing explanation

Fig. 1 is existing purification column structural representation.

Fig. 2 is the sectional view of Fig. 1.

Fig. 3 is the upper end, outer of the utility model ring-type trim ring and the interior structural representation along forming tilting plane between lower end.

Fig. 4 is the upper end, outer of the utility model ring-type trim ring and the interior structural representation along forming the cambered surface to lower recess between lower end.

In figure, 1 be ring-type trim ring, 2 be shell, 3 be filtering layer, 4 for liquid discharge pipe.

Embodiment

Embodiment 1.

As shown in Figure 3, for the purification column that body fluid trace dna extracts, comprise shell 2, be provided with filtering layer 3 in shell 2 lower inside, above filtering layer 3, be provided with ring-type trim ring 1, it is characterized in that: the upper end, outer of described ring-type trim ring 1 and interiorly form tilting plane between lower end; Liquid discharge pipe 4 is provided with in shell 2 lower end; The liquid such as elution reagent are discharged from liquid discharge pipe.

Described filtering layer 3 is sieve plate or filter paper.

Described shell 2 is column plastic casing.

Embodiment 2.

As shown in Figure 4, for the purification column that body fluid trace dna extracts, comprise shell 2, be provided with filtering layer 3 in shell 2 lower inside, above filtering layer 3, be provided with ring-type trim ring 1, it is characterized in that: the upper end, outer of described ring-type trim ring 1 and the interior cambered surface along forming between lower end to lower recess; Liquid discharge pipe 4 is provided with in shell 2 lower end; The liquid such as elution reagent are discharged from liquid discharge pipe.

Described filtering layer 3 is sieve plate or filter paper.

Described shell 2 is column plastic casing.

Claims (7)

1. the purification column extracted for body fluid trace dna, comprise shell (2), filtering layer (3) is provided with in shell (2) lower inside, be provided with ring-type trim ring (1) in filtering layer (3) top, it is characterized in that: the upper end, outer of described ring-type trim ring (1) and interiorly form high outside and low outside scarp between lower end.

2. a kind of purification column extracted for body fluid trace dna according to claim 1, is characterized in that: the upper end, outer of described ring-type trim ring (1) and interiorly form tilting plane between lower end.

3. a kind of purification column extracted for body fluid trace dna according to claim 1, is characterized in that: the upper end, outer of described ring-type trim ring (1) and the interior cambered surface along forming between lower end to lower recess.

4. a kind of purification column extracted for body fluid trace dna according to claim 1, is characterized in that: be provided with liquid discharge pipe (4) in shell (2) lower end.

5. a kind of purification column extracted for body fluid trace dna according to claim 1, is characterized in that: described filtering layer (3) is sieve plate.

6. a kind of purification column extracted for body fluid trace dna according to claim 1, is characterized in that: described filtering layer (3) is filter paper.

7. a kind of purification column extracted for body fluid trace dna according to claim 1, is characterized in that: described shell (2) is column plastic casing.