31 results about "Silver Staining Method" patented technology

The invention discloses a Coomassie brilliant blue dyeing method and its special gel stationary liquid and coloring agent. It includes the following steps: putting SDS-PAGE glue into the gel stationary liquid for 0.5-2.0h at normal temperature; adding the glue in acetate solution with 0.5-3.0% volume percent concentration to extend for 1.0-3.0h at normal temperature; adding it to the Coomassie brilliant blue coloring agent to dye for 8-24h at normal temperature; add it to decolorizing solution I to decolorize for 1-3h; then adding to decolorizing solution II to decolorize to clear. The invention can detect SDS-PAGE glue protein at nanogram.

The invention discloses a silver staining method of a polyacrylamide gel, which comprises the following steps of: firstly, putting an electrophoresed gel into a ceramic disc filled with distilled water; secondly, rinsing the gel by using the distilled water for two times; thirdly, adding a staining solution to carry out the solver staining; fourthly, re-rinsing the gel by using the distilled water for two times; fifthly, adding a developing solution to develop; and finally, using a digital camera to take pictures for the gel. By simplifying the silver staining steps, the silver staining method reduces the types of prepared solutions, simplifies a developing process, saves the dosage of a reagent decreasing the cost, and greatly shortens the developing time and improves the dye staining efficiency on the promise of guaranteeing obtaining an ideal target stripe. The obtained target section is clear enough for convenient genetype judgment and is particularly suitable to be used in processes of studying molecular markers and drawing genetic maps.

The invention discloses an ultra-fast silver staining test method of cane molecule tagged PAGE gel, which comprises the following steps: a modified polyacrylamide gel with the concentration of 5 percent is used for the electrophoresis detection of SSR and AFLP tagged PCR amplified primer; the gel after the electrophoresis is placed into a dyeing liquid to be dyed; the gel is taken out from the dyeing liquid, quickly rinsed, and placed into a developing solution to be developed; the developed gel is cleaned by distilled water for two times and dried, and tape reading is implemented; the ultra-fast silver staining test method combines four traditional steps of fixing, dyeing, developing and image fixing into two simplest steps, thus simultaneously reducing rinsing frequency and time, reducing the use of a fixing liquid, an image fixing liquid, acetic acid, absolute ethyl alcohol, sodium carbonate and sodium hyposulfite, not only greatly decreasing silver staining time and reagent cost, but also ensuring high-quality silver staining effect, and compared with other traditional or improved isotopic silver staining methods, the ultra-fast silver staining test method has larger time and cost advantages.

The invention discloses a method for molecular identification of Siniperca kneri and Siniperca scherzeri Steindachner and a kit used by the same. The method for molecular identification comprises the following steps: DNA extraction, DNA fragment amplification, electrophoresis of PCR products and identification by a silver-staining method. The kit contains a pair of SSR primers and a standard map. The method for molecular identification can improve the efficiency and accuracy of identification of Siniperca kneri and Siniperca scherzeri Steindachner, and is expected to solve the identification problem in the aquiculture of Siniperca kneri and Siniperca scherzeri Steindachner and stimulate the sound development of Siniperca chuatsi culture industry.

The invention discloses a rapid neuron staining method based on a Golgi silver staining method. The rapid neuron staining method based on the Golgi silver staining method is characterized in that according to mass volume ratio concentration, the aqueous solution with 5 % of potassium dichromate, the aqueous solution with 5 % of mercuric chloride and the aqueous solution with 5 % of potassium chromate are prepared into a Golgi silver staining solution in the volume ratio of 1:1:1, a brain tissue is soaked in the silver staining solution at 37 DEGCfor 36-48 hours, the soaked brain tissue is sliced on a vibrating slicer, tissue sections are arranged on a glass slide coated with gelatin, the tissue sections stay overnight, then conventional ammonia developing is conducted, gradient alcohol dehydration is conducted, the transparency process is conducted by xylene, finally, the tissue sections are sealed by netrual gum, and the tissue sections are observed through a microscope. According to the rapid neuron staining method based on the Golgi silver staining method, staining can be conducted on each encephalic region of the brain, the staining result is clear, details are obvious, and research staff can observe the neuron structure of the brain tissues conveniently, the convenient and fast means is provided for case analysis of nervous system lesions, and the rapid neuron staining method based on the Golgi silver staining method has significant meanings for the field of foundational research of neurology.

The invention discloses a polyacrylamide gel (PAG) solution for microsatellite marker (also known as simple sequence repeats or SSR) polymorphism detection. The polyacrylamide gel solution includes a polyacrylamide gel mother liquor, 5*TBE, a 10% APS and TEMED, diluting is carried out by adopting distilled water until the volume of the polyacrylamide gel mother liquor accounts for 21-22% of the total volume, and the amount of the 5*TBE is the same as that of the polyacrylamide gel mother liquor. When the polyacrylamide gel solution is used, PAG has less damage during an oscillation developing process, PAG splicing is avoided, the experimental error is reduced, and the accuracy of results is guaranteed.

The invention discloses a DNA silver staining method. The method comprises the following steps: (1) cutting off power supply after finishing electrophoresis, pouring a buffer solution out from an electrophoresis tank, then taking off a gel plate, putting the rubber plate into a porcelain dish filled with distilled water, and rinsing the gel plate by the distilled water for 2 to 3 times; (2) carrying out silver staining: adding a staining agent to carry out silver staining, shaking ceaselessly, staining for 8 to 12 minutes, and then rinsing by the distilled water for 2 to 3 times; (3) carrying out developing: adding a developing agent to carry out color development, shaking ceaselessly, and developing for 8 to 12 min; (4) rinsing by the distilled water for 2 to 3 times; and (5) carrying out gel shooting: spreading gelatin onto a X-ray film viewer to take a photo. The method integrates the fixing step and the staining step, adopts ethanol to fix, and overcomes the defects that the belt color is lighter when acetic acid is used for fixing, and the acetic acid has penetrating odor, causticity and so on in the prior sliver staining method. Additionally, the method does not need use ethanol and/ or acetic acid to stop developing color, omits the color development stopping step, and solves the problems of penetrating odor and causticity brought by the acetic acid and the like.

The invention discloses a DNA silver staining method in polyacrylamide gel (PAGE) electrophoresis, which comprises the steps of soaking polyacrylamide gel with 0.1% AgNO3 for 10-15min, washing with distilled water twice, developing with 0.04% Na2CO3, 2mL of 37% HCOH/L and 2% mixed solution for 5-6min, and flushing twice with distilled water to complete the whole dyeing process. The conventional DNA silver staining method needs 5-6 chemical reagents, prepares 4 solutions and spends 40-60min to complete the dyeing step, but the method only needs 4 chemical reagents, prepares 2 solutions and spends 20min to reach the result the same as that of the conventional dyeing method, and in denaturing polyacrylamide electrophoresis (PAGE), the dyeing method can reduce the amount of DNA to 0.44ng; and in non-denaturing polyacrylamide electrophoresis (PAGE), the amount of DNA can be reduced to 3.5ng. Compared with the conventional dyeing method, the method of the invention saves the time, reduces the cost, and increases the detection sensitivity.

The invention relates to a method for dyeing DNA (Deoxyribose Nucleic Acid) on polyacrylamide gel. The method is a safe and environment-friendly method for dyeing acidic silver ion solution. The method sequentially comprises the steps of fixing, primary washing, impregnating, secondary washing, developing, and developing reaction ending. In addition, the invention further relates to a method for detecting the DNA on the polyacrylamide gel, and a kit for the methods.

The invention discloses an optimized rice indica and japonica performance detection system. The optimized rice indica and japonica performance detection system comprises a molecular marker system, an amplification system, an electrophoresis system and a gel imaging system, wherein the molecular marker system is used for molecular marking of a group of indica and japonica high in specificity, the amplification system is used for PCR (polymerase chain reaction) amplification, the electrophoresis system is used for PAGE (polyacrylamide gel electrophoresis), and the gel imaging system is used for imaging detection of banding patterns. The optimized rice indica and japonica performance detection system has the advantages that the detected banding patterns are clear and obvious, and adopting a GoldView aqueous solution for staining substitutes for a traditional PAGE gel silver staining method, so that an operation process is simpler and more convenient, and the amplification efficiency and the detection efficiency are both improved. The invention further provides a rice indica and japonica performance differentiating method. A designed indica and japonica performance differentiating marker is high in specificity, only one indica or japonica specific band pattern occurs, and the occurrence frequency of the specific band pattern in an indica and japonica group is above 70.0%. The indica and japonica performance is measured through the indica and japonica degree, and accordingly comparison results are more objective and comprehensive.

The present invention relates to a method for molecular auxiliary selection of Dian-type hybrid rice restoring line, belonging to the field of breeding technology. Said method includes the following steps: sowing and transplanting the material to be identified, after the material is transplanted for 15-40 days, taking leaf, extracting total DNA of rice; determining sample DNA concentration and quantitatively diluting DNA to 25 ng/ul; then using micro-satellite marker OSR33 and RM228 which are positioned in tenth chromosome of rice and tightly linked with Dian-type restoring gene as primer, making PCR reaction; making poly-acrylamide gel electrophoresis, utilizing silver staining method to detect PCR product and electrophoresis result; then judging that the tested material has restoring gene or not and can be used as restoring line of Dian-type hybrid rice or not. Said invention can quickly and effectively screen restoring line of Dian-type hybrid rice.

The invention discloses a method for growing DNA at the 3'-OH terminal of RNA based on TdT and free of template dependence. The method mainly comprises the following steps: with miRNA122 and DNA122 asprimers, adding dNTPs, 1*CoCl2, 1*RB10* and TdT to form a 20-microliter reaction system; adjusting reaction conditions including reaction time, reaction concentration, reaction raw materials and reaction substrates; verifying a RNA extension effect through polyacrylamide gel electrophoresis and a silver staining method; and verifying the difference of extension effects of DNA and RNA under the action of the TdT through a fluorescence method. The invention relates to the novel application of TdT; and a series of high-sensitivity RNA detection platforms can be established by utilizing the new application, and a new detection direction is provided for miRNAs such as tumor markers in the future.

The invention provides a kit and an identification method for molecules of five Sinipercinae fishes. The kit comprises three pairs of SSR primers and standard maps. The identification method for molecules of five Sinipercinae fishes by using the kit comprises the following steps: 1) DNA extraction: a step of acquiring the fin ray or muscle tissue of a to-be-tested Siniperca chuatsi sample, extracting genome DNA and measuring the purity and concentration of the genome DNA; 2) DNA fragment amplification: a step of amplifying the genome DNA of the to-be-tested sample by using a polymerase chain reaction; 3) electrophoresis and silver staining identification of PCR products: a step of carrying out electrophoretic separation on the PCR products by using nondenaturing polyacrylamide gel, then carrying out staining by using a conventional silver staining method and taking photos to record electrophoresis results; and 4) comparison with the standard maps. The kit and the identification method provided by the invention can substantially improve efficiency and accuracy of identification of important economic and rare varieties of Sinipercinae and are beneficial for protection of genetic resources of Sinipercinae fishes and sustained sound development of the breeding industry of Siniperca chuatsi.

The invention discloses a method for screening the allelotype of an immune related gene DQA1 Exon 2-(A-C) of goat. A primer is designed according to the sequence of exon-2 of a DQA1 gene, the genome DNA of white goat is used as a template to carry out PCR (Polymerase Chain Reaction) amplification, purification sequencing is carried out on the PCR product, and the polymorphic site of the exon-2 of DQA1 gene is screened preliminarily; a target fragment is subjected to PCR amplification by using DNA of the white goat genome as a template, and the PCR product is treated by a denaturing agent, and the denatured amplification fragment is subjected to polyacrylamidedel gel electrophoresis detection; rubber is dyed by applying a silver staining method, and partial allelotype of the exon-2 of the DQA1 gene of white goat from Guizhou is determined according to the result of polyacrylamidedel gel electrophoresis. A convenient, quick and accurate method is provided to PCR-SSCP screening and type determining of exon-2 polymorphism of DQA1 gene of different varieties of goat, and can be used for detecting partial allele of exon2 of goat immune related DQA1 gene, so that the problem of difficult type determination on highly polymorphic genes in the existing PCR-SSCP technology can be solved.

The invention provides a quantitative protein silver staining method of freshwater ciliates. The method includes sample fixation, filtration and concentration, agar embedding, agar solidification, protein silver staining and dehydrating and mounting. According to the method, a staining effect is better, slices produced in unit time are more, an overall staining effect is uniform, and mounting is easy. The method is more suitable for protein silver staining technology of qualitative and quantitative study of planktonic ciliates in typical eutrophic lakes of China.