正向引物primer-f:cagcaatgggtaggaga; 反向引物primer-r:cttctcagggccccag。 一种用于检测tnf-a基因snp位点rs1799724基因型的试剂盒,所述的试剂盒包括有上述的引物对和特异探针,所述的特异探针序列probe-c:ccccttaacgaagacag,序列5’段使用fam标记,3’端用mgb标记;特异探针序列probe-t:ccccttaatgaagacag,序列5’...
根据NCBI数据库中小鼠TNF-α和GAPDH的基因序列,应用Primer Premier 5.0软件设计正、反义引物和探针,引物和探针均由上海生工公司合成和修饰,见表1。 1.5 实时荧光PCR 总反应体系为25μL,包括cDNA模板2μL,50 mmo1/L MgC12溶液1.5μL,IMMOLASE DNA热启动酶0.2μL,100μmo1/L TNF-α及GAPDH正、反义引物和探...
Genomic DNA was obtained from oral swabs of all subjects and amplified using the polymerase chain reaction (PCR) with specific primers flanking the locus −308 of TNF-α. PCR products were submitted to restriction endonuclease digestion and analysed by polyacrylamide gel electrophoresis to ...
The primers used for the real-time RT–PCR were purchased from Takara. For laser microdissection-based RT–PCR, epithelial cells and stromal cells were separately collected from frozen sections of Gan mouse tumors (n ¼ 4) using laser microdissection LMD7000 (Leica Microsystems, Wetzler, ...
The primer pairs specific for various genes used in our experiments are shown in Supplementary Table 1. A triplicate sample was analysed in each group. siRNA and shRNA interference. siRNA targeting human TNFR1, TNFR2, VEGFR3, scrambled siRNA and transfecting medium were purchased from Dharmacon ...
5’-TGGTATGAAATGGCAAATCG-3’,特异性扩增片段为329bp);IL-1B引物(Forward Primer:5’-CTCATTGTGGCTGTGGAGAA-3’,Reverse Primer:5’-GGGATTTTGTCGTTGCTTGT-3’,特异性扩增片段为323bp);B-actin引物(Forward Primer:5’-GCCGTCTTCCCCTCCATCGTG-3’,Revere Primer:5’-TAC GACCAGAGGCATACAGGGACAAC-3’...
为得到含有9号外显子的产物,需将primerexon7f和primerexon7r替换为特异性引物primerexon9f和primerexon9r,阳性对照则需替换为t-9质粒,其它条件不变。扩增的条件为:95℃预变性5min,再进行35个循环,循环条件为95℃变性30s,51℃退火30s,72℃延伸30s;最后72℃延伸5min。得到的pcr产物进行凝胶电泳,结果如图1所示。
ainsiu’une raideur avec déroui l lage matinal Le syndrome inflammatoire général peut’exprimer par une fatigue,une inappétence,une perte de poids et de la fièvreEnsuite, les patients présentent des polyarthrites, puis des dommages articulairesvec destructions et déformations irréversibles...
Genomic DNA was extracted; PCR–RFLP was performed usingTNF-αprimers specific to detect the presence of SNPs. Various bioinformatics tools such as STRING software was used to determine its network with other associated genes. The PCR–RFLP studies showed that among the 238G/A types the GG ...
primer: 5 0 -TCCT-CCCTGCTCCGATTCCG-3 0 . Cycling was carried out for 1 cycle at 94 °C for 3 min, 35 cycles for 1 min each at 94 °C, 60 °C and 72 °C; and 1 cycle at 72 °C for 1 min. Digestion of PCR products with NcoI yielded 87 + 20-bp fragments (allele 1: ...