Rapid and low-cost method for separating PCR template from chicken blood

Abstract

The invention discloses a rapid and low-cost method for separating a PCR template from chicken blood. The rapid and low-cost method for separating the PCR template from the chicken blood comprises thesteps that A, blood corpuscle nucleuses are separated from a blood sample, specifically, a fresh and frozen anticoagulant chicken blood sample is added into a 1.5 mL EP pipe, and 4000rpm centrifugation is carried out for one minute and supernatant is removed; and 500 [mu]l sterile water is added into the pipe to stand for one minute, 9500rpm centrifugation is carried out for three minutes, and the supernatant is abandoned; B, the corpuscle nucleuse dissociation is carried out, specifically, a 3-4% chelated type ion exchange resin solution shaken up by the 200 [mu]l sterile water is added intosediment by using a spearhead with the cut tip part, after mixing, water bath at 55 DEG C-57 DEG C is carried out for 30 minutes, then, a dry heat meter is placed in, and heating is carried out at 100 DEG C for 8 minutes; and C, PCR template separation is carried out, specifically, 9500rpm centrifugation is carried out for two minutes, and the supernatant is taken as a PCR template. Compared witha template obtained by using an existing traditional DNA extraction method, the PCR template obtained by using the rapid and low-cost method for separating the PCR template from the chicken blood hassame effects for PCR technology detecting, the operation is simpler and easier to operate, the time-consuming is short (within one hour), the cost is low (about 0.31 yuan / sample), and the PCR template is suitable for various rapid DNA molecular detection experiments carried out by using PCR technology.

Description

technical field [0001] The invention belongs to the technical field of rapid nucleic acid extraction, and in particular relates to a method for separating PCR template (DNA) from chicken blood rapidly and at low cost. detection experiment. Background technique [0002] With the rapid development of molecular biology technology, various DNA molecular detection technologies have been gradually applied to the fields of medicine, agriculture, environment and biology. At present, in the field of agriculture, PCR amplification technology has been widely used in breeding or disease diagnosis work, and can detect the associated genes of specific diseases and related traits. In some of these works, rapid identification or diagnosis is required, and some experiments require a large number of samples to be detected; however, the time and cost of template DNA isolation in PCR detection severely restrict the widespread use of such techniques. Therefore, how to separate PCR templates qui...

AI Technical Summary

Problems solved by technology

In the current PCR detection practice, the traditional DNA extraction method is time-consuming, which is not enough to meet the needs of more efficient and rapid acquisition of PCR templates

In order to isolate PCR templates (DNA) from chicken blood in large quantities more quickly and at low cost, the applicant, on the basis of referring to and improving previous work, screened out a chelating ion exchange resin to lyse the nucleus and established a A method that can obtain PCR templates from chicken blood quickly and at low cost. The obtained PCR templates (DNA) have the same efficacy as the templates obtained by existing traditional DNA extraction methods for PCR technology detection. No one has been found after searching Rapid, low-cost method for isolating PCR template (DNA) from chicken blood is disclosed or used

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