48 results about "Trizol" patented technology

The invention provides a method for separating exosomes from animal plasma and for detecting purity. The method mainly comprises the following steps: first, thinning the plasma by virtue of a PBS (phosphate buffer solution) at the ratio of 1 to 1; removing cell components and cell debris in the plasma through centrifuging; precipitating the exosomes through ultrahigh-speed centrifugation; suspending the exosomes by virtue of 250[mu]l of the PBS solution; extracting total RNA in an exosome suspension by virtue of Trizol LS; and conducting poly (A) reversing and related gene quantification on the extracted total RNA, and detecting the purity. The method disclosed by the invention is conducive to researches on plasma exosome contents and exosome related functions in the future.

The invention discloses a kit used for detecting relative expression of the AML1-ETO fusion gene. The kit comprises erythrocyte lysate, TRIzol, chloroform, anhydrous ethanol, a ReverTra Ace qPCR RT kit, a detection system PCR reaction solution, a positive control and a negative control, and is characterized in that the detection system PCR reaction solution comprises THUNDERBIRD qPCR MIX, the upstream and downstream primers of AML1-ETO-F and AML1-ETO-R and the probe of AML1-ETO-Probe used for detecting target genes, and the primers of ab1-F and ab1-R and the probe of ab1-Probe used for detecting the internal control gene Ab1. The kit provided in the invention can detect the expression level of the AML1-ETO fusion gene in patients with acute myeloid leukemia (AML) and can effectively save detection time and improve detection precision.

The invention provides a method for separating exosomes from animal plasma and for detecting purity. The method mainly comprises the following steps: first, thinning the plasma by virtue of a PBS (phosphate buffer solution) at the ratio of 1 to 1; removing cell components and cell debris in the plasma through centrifuging; precipitating the exosomes through ultrahigh-speed centrifugation; suspending the exosomes by virtue of 250[mu]l of the PBS solution; extracting total RNA in an exosome suspension by virtue of Trizol LS; and conducting poly (A) reversing and related gene quantification on the extracted total RNA, and detecting the purity. The method disclosed by the invention is conducive to researches on plasma exosome contents and exosome related functions in the future.

The prostate cancerometastasis agent box uses a timely RT-PCR technique, starting from the erythrocytoschisis to separate the surrounding blood individual cell, extracting the overall RNA of each cell through Trizol, reversely transcribing the single cell RNA into CDNA No. 1 chain, achieving timely quantitative RT-PRC inspection through TaqMan detective needle. It improves the sensitivity and astopic feature of PSA and PSMAmRNA inspection, effectively getting over the defects of normal RT-PCR extension, with the precision could detecting one prostate within 107 single cells. This inspection method, design lead, detective needle and clinical sample inspection result provides reliable reference for the development of scheduled quantitative PCR inspection agent box.

The invention relates to an animal genetic engineering interferon compound preparation, the production method and the clinical application. The animal genetic engineering interferon compound preparation is prepared by extracting the animal peripheral blood or spleen lymphocytes, culturing, inducing by in vitro inductor, extracting the cell total RNA by Trizol, designing the specific primer, cloning the interferon gene by RT-PCR technology and connecting to the pGEM-T carrier by T-A strategy; the primer is designed again, the both ends are added with different restriction endonucleases digestion sequences, initiation codon ATG and termination codon TAA, leading peptide sequences are removed, the PCR amplification is carried out by taking the recombination T carrier as the template, so as to obtain the expression fragment of the animal interferon gene; the expression fragment is carried out with the rare codon mutation, and gel recovery target fragment is connected on the carrier of pFastBacDual with the same double restriction endonucleases digestion after double restriction endonucleases digestion; the interferon-Alpha of an animal is cloned to multiple cloning sites under the control of Polyhedrin promoter, the interferon-Gamma of the same species animal is cloned to multiple cloning sites under the control of p10 promoter; the constructed carrier of pFastBacDual plus interferon Alpha plus interferon Gamma are transfected to DH10BacE.coli to carry out recombination; the constructed recombinant bacmid is extracted and purified after the blue white screening and the PCR identification, and the transfection of SF9 insect cell is carried out by a liposome method; the expression product is carried out with identification and purification; the animal genetic engineering interferon compound preparation is carried out with anti-virus activity detection and the evaluation of the clinical application effects.

The SCNN1 primer for rapidly diagnosing clear cell nuclear cell carcinoma provided by the invention comprises an internal reference primer pair which is GAPDH, mRNA sequence forward primers and reverse primers of SCNN1A genes with nucleotide sequences shown in SEQ ID NO: 1 and SEQ ID NO: 2, mRNA sequence forward primers and reverse primers of SCNN1B genes with nucleotide sequences shown in SEQ IDNO: 3 and SEQ ID NO: 4, and mRNA sequence forward primers and reverse primers of SCNN1G genes with nucleotide sequences shown in SEQ ID NO: 5 and SEQ ID NO: 6, which take human GAPDH as an internal reference control, and forward primers and reverse primers, having nucleotide sequences shown as SEQ ID NO: 7 and SEQ ID NO: 8. A corresponding kit is also prepared. The kit comprises the following reagents: a tissue lysis solution Trizol, an SYBRGreen Master Mix reaction solution, a reverse transcription reaction solution, a negative control and a positive control. The primer and the kit can eliminate false negative and false positive of clinical sample detection, and are safe, reliable, high in accuracy and suitable for clinical popularization.

The invention belongs to the field of biotechnology, and specifically relates to an RNA extraction device and method, comprising a piston syringe, a plastic centrifuge tube, a centrifugal column arranged in the plastic centrifuge tube, a mixing centrifuge tube, the mixing centrifuge tube includes a tube body and a tube cover, A through hole is arranged on the tube cover, a silica gel column is arranged in the through hole, a top separation layer is arranged on the upper part of the inner cavity of the tube, and a middle separation layer is arranged on the middle and lower part, and the top separation layer and the middle separation layer separate the inside of the tube body. The chamber is divided into an upper chamber, a middle chamber and a lower chamber. Grinding balls are pre-placed in the upper chamber, TRIzol is pre-placed in the middle chamber, and chloroform is pre-placed in the lower chamber. In the process of extracting RNA in the present invention, the operator does not need to directly absorb and contact the organic solvent, and no toxic volatiles are released into the atmosphere during the operation, which protects the health of the experimenters; at the same time, the present invention realizes the detection of bulk tissue samples through grinding balls. Mechanical cutting and grinding of difficult-to-lyse tissue samples to release RNA more quickly and fully.