36results about How to "Solve the current situation of no internal reference genes" patented technology

The invention provides an internal reference gene of chives and its screening method and application, belonging to the field of molecular biology. Select 13 candidate internal reference genes Ac18S, Af18s, Af28s, AcActin, AfActin, AfTUA, AfTUB, AsSAND, AfMYH, AfGAPDH, AfUBQ, AfEF1a, AfCYC, and use real-time fluorescent quantitative RT-qPCR technology to express their expression levels under high temperature stress The stability of the 13 internal reference genes was evaluated in combination with GeNorm, NormFinder and BestKeeper software, and finally a comprehensive analysis was conducted to obtain the most suitable internal reference gene for chives under high temperature stress as AfActin, which provided a basis for subsequent scientific research on gene expression in chives. refer to.

The invention provides a passion flower reference gene PeNADP and a screening method and an application thereof, which belong to the field of molecular biology. The RNA-Seq sequencing data is used foranalyzing moderate or highly stable genes in the expression quantity to serve as candidate reference genes; according to the expression quantity value of the PeNADP, two genes with the highest expression quantity and two medium-high expression genes are selected, the primer is designed, verification is carried out on mRNA of different tissue parts of the passion flower variety with obvious phenotypic difference, the expression stability of each candidate reference gene is evaluated by utilizing software, and finally the proper passion flower reference gene PeNADP is screened out. The reference gene PeNADP can be stably expressed in stems, leaves, flowers and fruits of Tainung No.1 and gold berry No.2., and reference is provided for subsequent scientific research related to passion flowergene expression.

The invention discloses internal reference genes of different tissues of loofah, which are PP2A and / or β-ACTIN. The sequence of the internal reference gene PP2A is shown in SEQ ID No.1; the sequence of the internal reference gene β-ACTIN is shown in SEQ ID No. 2. The internal reference genes PP2A and β-ACTIN of the present invention have higher stability than other internal reference genes, and these two internal reference genes are screened out, and are the most stable internal reference genes in different tissues of loofah, and the data are true and reliable. Given the fact that there is no internal reference gene in loofah gene expression analysis, it can also improve detection efficiency and detection results, and has high practical value. It provides strong support for obtaining accurate quantitative data of functional gene expression in different tissues of loofah, and provides a basis for gene function of loofah. Research provides a theoretical basis.

The invention discloses a fluorescent quantitative internal reference gene and its primers and applications under the stress of Changshan salt in Haizhou. The internal reference genes are UBC‑E2, RPL and MDH, wherein the gene sequence of the MDH gene is shown in SEQ ID NO.10, and the RPL gene The gene sequence of the UBC‑E2 gene is shown in SEQ ID NO.3, and the gene sequence of the UBC‑E2 gene is shown in SEQ ID NO.12. This application screens out candidate genes with relatively stable expression based on the existing published genes, evaluates their stability through software, discloses the most stable internal reference genes suitable for Haizhou Changshan salt stress conditions, and uses these genes as a basis to design Real-time fluorescence quantitative PCR primers fill the status quo of no internal reference genes under the salt stress conditions of Changshan Mountains in Haizhou, provide strong support for the accurate quantification of related functional genes under the conditions of salt stress in Changshan Mountains in Haizhou, and can improve the stability, repeatability and reliability of research sex.

The invention provides a passion flower reference gene as well as a screening method and an application thereof, which belong to the field of molecular biology. The RNA-Seq sequencing data is used foranalyzing genes with high and stable expression quantity to serve as candidate reference genes, five genes with high expression quantity (FPKM is approximately equal to 850) are selected, primer designing is carried out, verification is carried out in mRNA of different tissue parts of a passion flower variety with obvious phenotypic difference, the expression stability of each candidate referencegene is evaluated by utilizing software, and finally a proper passion flower reference gene C21209 is screened out. The C21209 can be stably expressed in stems, leaves, flowers and fruits of TainungNo.1 and golden fruit No.2, and reference is provided for subsequent scientific research related to passion flower gene expression.

The invention discloses a fluorescent quantitative internal reference gene under salt stress of Haizhou Changshan Mountain and primers and applications thereof, wherein the internal reference gene isUBC-E2, RPL and MDH, wherein the gene sequence of MDH gene is shown in SEQ ID NO:10, the gene sequence of RPL gene is shown in SEQ ID NO:3, the gene sequence of UBC-E2 gene is shown in SEQ ID NO.12. The present application screens out candidate genes with relatively stable expression according to the genes disclosed in the prior art, its stability is evaluated by software, a most stable internal reference gene suitable for expression under salt stress condition in Haizhou Changshan mountain is disclosed, Based on these genes, real-time fluorescence quantitative PCR primers were designed, whichfilled the situation the there were no internal reference genes under salt stress in Haizhou Changshan Mountain, and provided a powerful support for the precise quantification of related functional genes under salt stress in Haizhou Changshan Mountain, and could improve the stability, repeatability and reliability of the research.

A fluorescence quantitative internal reference gene suitable for Haizhou Changshan flower, its primer and the application thereof are disclosed, wherein the internal reference gene is Actin and AP-2 gene, wherein the gene sequence of the Actin gene is as shown in SEQ ID NO: 1, AP-2 gene is shown in SEQ ID NO: 13. The present application screens out candidate genes with relatively stable expressionaccording to the genes disclosed in the prior art, its stability is evaluated by software, A fluorescence quantitative internal reference gene suitable for Haizhou Changshan flower was obtained, Based on these genes, real-time fluorescence quantitative PCR primers were designed, which filled the situation that there were no internal reference genes in different flowering stages of Haizhou Changshan from different provenances, and provided a powerful support for accurate quantification of related functional genes in different flowering stages of Haizhou Changshan from different provenances, and could improve the stability, repeatability and reliability of the research.

The invention provides primers for screening internal reference genes under low temperature stress in lotus mist and applications thereof. The primers are: forward primer 5'-CCTGGCCTCTATGATCTTATCC-3', reverse primer 5'-CAGGCATCATGTGCTTTG-3'. The present invention uses qRT-PCR to detect the expression levels of 7 candidate internal reference genes ACT-7, GAPDH, UBQ, α-TUB, CYP20-1, EF-2 and APRT-5 under the development of lotus mist fruit and low temperature stress, through geNorm , NormFinder, BestKeeper and RefFinder software for gene stability evaluation, the results show that the expression of the most stable internal reference gene CYP20-1 under low temperature stress, this not only solves the current situation that there is no internal reference gene in the existing lotus mist quantitative PCR detection, but also provides a basis for the follow-up Laying the groundwork for gene expression work.

The invention provides a passion flower reference gene PeGBP and a screening method and an application thereof, which belong to the field of molecular biology. The RNA-Seq sequencing data is used foranalyzing moderate or highly stable genes in the expression quantity to serve as candidate reference genes; according to the expression quantity value of PeGBP, two genes with the highest expression quantity and two medium-high expression genes are selected, the primers are designed, verification is carried out in mRNA of different tissue parts of the passion flower variety with obvious phenotypicdifference, the expression stability of each candidate reference gene is evaluated by utilizing software, and finally a proper passion flower reference gene PeGBP is screened out. The reference genePeGBP can be stably expressed in stems, leaves, flowers and fruits of Tainung No.1 and gold berry No.2., and reference is provided for subsequent scientific research related to passion flower gene expression.

The invention provides a passion flower reference gene as well as a screening method and an application thereof, which belongs to the field of molecular biology. The RNA-Seq sequencing data is used for analyzing stable genes with the medium expression level to serve as the candidate reference genes; five genes with medium expression quantity (FPKM is approximately equal to 25) are selected, primerdesigning is carried out, verifying is carried out in mRNA of different tissue parts of passion flower varieties with obvious phenotypic difference, the expression stability of each candidate reference gene is estimated by utilizing software, and finally the proper passion flower reference gene C27182 is screened out. The C27182 can be stably expressed in stems, leaves, flowers and fruits of Tainung No.1 and golden fruit No.2. The reference is provided for subsequent scientific research related to passion flower gene expression.

The present invention discloses a fluorescent quantitative internal reference gene suitable for different development stages of Changshan leaves in Haizhou and its primers and applications. The internal reference genes are Actin, RAN and SAND genes, wherein the gene sequence of the Actin gene is shown in SEQ ID NO.1, The gene sequence of the RAN gene is shown in SEQ ID NO.6, and the gene sequence of the SAND gene is shown in SEQ ID NO.8. Based on the existing published genes, this application screens out candidate genes with relatively stable expression, evaluates their stability through software, obtains fluorescent quantitative internal reference genes suitable for different developmental stages of Changshan leaves in Haizhou, and uses these genes as a basis to design a real-time Fluorescent quantitative PCR primers fill in the current situation that there are no internal reference genes in different leaf stages of Haizhou Changshan in different provenances, and provide strong support for the accurate quantification of related functional genes in different leaf stages of Haizhou Changshan in different provenances, which can improve the research efficiency. Stability, repeatability and reliability.

The invention discloses a fluorescent quantitative internal reference gene and its special primers and applications under drought stress in Changshan, Haizhou. The fluorescent quantitative internal reference genes are Actin, AP‑2 and RAN genes, wherein the gene sequence of the Actin gene is as shown in SEQ ID NO.1 The gene sequence of the AP‑2 gene is shown in SEQ ID NO.13, and the gene sequence of the RAN gene is shown in SEQ ID NO.6. The present invention screens candidate genes with relatively stable expression from transcriptome data and internal reference genes studied by previous studies, and evaluates their stability through software, and discloses three internal reference genes suitable for the most stable expression under drought stress in Changshan, Haizhou , and based on these genes, real-time fluorescent quantitative PCR primers were designed to fill the status quo of no internal reference genes under drought stress in Changshan, Haizhou, and provide strong support for the accurate quantification of related functional genes under drought stress in Changshan, Haizhou, which can improve the stability of research repeatability, repeatability and reliability.

The present invention discloses fluorescent quantitative internal reference genes of different tissues in Changshan, Haizhou, primers and applications thereof. The internal reference genes are Actin, RPL, RAN, AP‑2 and HSP70 genes, wherein the gene sequence of the Actin gene is as shown in SEQ ID NO.1 As shown, the gene sequence of the RPL gene is shown in SEQ ID NO.3, the gene sequence of the RAN gene is shown in SEQ ID NO.6, the gene sequence of the AP-2 gene is shown in SEQ ID NO.13, and the gene sequence of the HSP70 gene is shown in SEQ ID NO.13. The gene sequence is shown in SEQ ID NO.14. The present invention designs an internal reference gene combination suitable for different tissues in Changshan, Haizhou, reveals the sequence of the internal reference gene, and designs real-time fluorescent quantitative primers accordingly, which not only solves the current situation that there is no internal reference gene in the existing detection of Changshan, Haizhou, but also The designed real-time fluorescent quantitative primers have strong primer specificity when used for gene expression analysis in Haizhou Changshan tissues, which can greatly improve the detection efficiency of real-time fluorescent quantitative detection of Haizhou Changshan genes, and improve the credibility of the detection results Spend.

The invention provides a Japanese maple reference gene, and also discloses a real-time fluorescence quantification PCR primer by using the Japanese maple reference gene as a basic design. The currentsituation that existing Japanese maple quantification PCR detection is free of a reference gene is solved; moreover, when the designed real-time fluorescence quantification PCR primer is used for analysis of Japanese maple reference gene expression, the stability, reliability and repeatability of analysis and research of Japanese maple gene expression can be improved; and moreover, the designed real-time fluorescence quantification PCR primer is applicable for related gene quantification expression research of different leaf stages and different leaf color varieties of Japanese maple, so thatthe accuracy of obtained data can be obviously improved, and technical support is provided for reducing the error of different samples, standardizing total RNA quantity and guaranteeing the reliability of data.

The invention provides a passion flower reference gene PeNADP and a screening method and an application thereof, which belong to the field of molecular biology. The RNA-Seq sequencing data is used foranalyzing moderate or highly stable genes in the expression quantity to serve as candidate reference genes; according to the expression quantity value of the PeNADP, two genes with the highest expression quantity and two medium-high expression genes are selected, the primer is designed, verification is carried out on mRNA of different tissue parts of the passion flower variety with obvious phenotypic difference, the expression stability of each candidate reference gene is evaluated by utilizing software, and finally the proper passion flower reference gene PeNADP is screened out. The reference gene PeNADP can be stably expressed in stems, leaves, flowers and fruits of Tainung No.1 and gold berry No.2., and reference is provided for subsequent scientific research related to passion flowergene expression.

The invention discloses an internal reference gene of purple cabbage, the base sequence of which is shown in SEQ ID NO:1. The invention discloses a purple cabbage internal reference gene amplification primer pair. The amplification primer pair is the base sequence shown in SEQ ID NO: 2 and 3. The invention discloses a pair of real-time fluorescent quantitative primers for internal reference genes of purple cabbage. The real-time fluorescent quantitative primer pair is: the base sequences shown in SEQ ID NO: 4 and 5. The invention discloses a kit for detecting the relative expression level of functional genes of purple cabbage, which comprises a pair of real-time fluorescent quantitative primers. The invention discloses the use of an internal reference gene or a real-time fluorescent quantitative primer for the detection of the relative expression levels of purple cabbage functional genes in different development stages of purple cabbage and in different varieties of purple cabbage. The invention solves the current situation that there is no internal reference gene in the existing quantitative PCR detection of purple cabbage, and improves the stability and reliability of gene expression analysis in detection of purple cabbage by using real-time fluorescent quantitative PCR technology.

The invention discloses a fluorescent quantitative internal reference gene and its special primers and applications under waterlogging stress in Changshan, Haizhou. The fluorescent quantitative internal reference genes are MDH, UBC‑E2 and UBQ, wherein the gene sequence of the MDH gene is as shown in SEQ ID NO.10 The gene sequence of the UBC-E2 gene is shown in SEQ ID NO.12, and the gene sequence of the UBQ gene is shown in SEQ ID NO.16. The present invention screens candidate genes with relatively stable expression based on the existing published genes, evaluates their stability through software, and obtains three internal reference genes suitable for the most stable expression under the waterlogging stress in Changshan, Haizhou, and uses these genes as a basis to design The real-time fluorescent quantitative PCR primers have filled the current situation that there is no internal reference gene under the waterlogging stress of Changshan in Haizhou, and provided strong support for the accurate quantification of related functional genes under the waterlogging stress of Changshan in Haizhou, which can improve the stability and repeatability of the research and reliability.

The invention discloses internal reference genes suitable for the expression research of lotus flood stress genes. The internal reference genes include: lotus actin gene and ribosomal protein 18sRNA gene. The present invention discloses two internal reference genes for studying the expression of water-flooding stress genes in lotus, and discloses real-time fluorescent quantitative PCR primers designed based on these two genes, solving the problem that there is no internal reference gene in the study of water-flooding stress gene expression in lotus The current situation of the current situation, the internal reference gene of the present invention can be stably expressed when the lotus responds to flooding stress, and the designed real-time fluorescent quantitative PCR primers are used for gene expression analysis when the lotus responds to flooding stress, which can improve the stability, reliability and reliability of the research. repeatability.

The invention provides a primer pair for screening reference genes of wax apple fruit development process and application thereof. The primers comprise an ACT-7 forward primer 5'-CGTTTGGCTCACTCCATACA-3', a reverse primer 5'-AGGTCTTTGTTGGTGAGCTATT-3'; an alpha-TUB forward primer 5'-TCCTTTAAGCCTATGTCGTAACC-3', and a reverse primer 5'-R: TCCGCAGCTTTCTTCTTT-3'. With the utilization of a qRT-PCR detection technology and through geNorm, NormFinder, BestKeeper and RefFinder software for gene stability evaluation, results show that reference genes which are most stably expressed during pulp and pericarp development process are respectively ACT-7 and alpha-TUB. The present situation that existing wax apple quantitative PCR detection has no reference gene is solved, and a foundation is also laid for subsequent gene expression work.