AmplideX® PCR/CE FMR1 Reagents
AmplideX PCR/CE FMR1 Reagents* are market-leading research tools for the detection of CGG repeats in the fragile X mental retardation (FMR1) gene. These reagents provide a PCR-only approach based on Triplet Repeat Primed PCR (TP-PCR) design to reliably amplify and detect all alleles including Full Mutations.
Features & Benefits
AmplideX PCR/CE FMR1 Reagents* have created an easy-to-use, accessible, high performance method for laboratories to reliably analyze CGG repeats and detect interrupting AGG sequences in the FMR1 gene.
Reduced ComplexityEase-of-analysis of the FMR1 gene has been simplified through:
- Implementation of proprietary PCR solution for amplifying GC-rich regions
- Automation of result calling using AmplideX PCR/CE FMR1 Reporter*
Optimized WorkflowValuable operator hands-on time has been significantly reduced through:
- Direct injection of PCR products (no PCR clean up) in to Capillary Electrophoresis platforms
- Decreased need for Southern blot analysis (up to 50 fold)
- End-to-end solution for FMR1 analysis including all necessary reagents and software
Quality PerformancePerforming FMR1 Analysis with Greater Sensitivity and Accuracy:
- Detection of all allele expansions, including low abundance full mutation size mosaics with up to at least 1300 CGG repeats
- Up to 875 fold more sensitive than Southern blot1
- Resolution of female homozygous and heterozygous samples and indication of interrupting AGG sequences
- Proven performance as indicated by more than 30 peer reviewed publications
*For Research Use Only. Not for use in diagnostic procedures.
Product Description
Analytical Characteristics of AmplideX PCR/CE FMR1 Reagents*:
- Detects all alleles including low abundance full mutations (Figure 1)
- Accurately sizes any repeat up to 200 CGG repeats (Figure 2)
- Resolves female zygosity (Figure 3)
- Detects presence of AGG interruptions (Figure 4)
Figure 1: Amplification of Asuragen’s Methylation and Sensitivity Control which has a 5% full mutation in a background of 95% Normal
Figure 2. Female premutation sample with accurate sizing of Normal (30 CGG) and Pre mutation allele (56 CGG)
Figure 3: The difference in the “stutter” peak patterns of homozygous and heterozygous female provides a clear resolution of zygosity
Figure 4. Female Full Mutation sample with AGG interruptions as indicated by sudden decrease in peak heights of the “stutter” peak profile
Ordering
Product Name | Number of Reactions | Catalog Number |
---|---|---|
AmplideX PCR/CE FMR1 Control | 24 UL | 49513 |
AmplideX mPCR FMR1 Control | 24 UL | 49514 |
AmplideX PCR/CE FMR1 Reagents | 100 | 49402 |
AmplideX mPCR FMR1 Kit | 24 | 49442 |
AmplideX PCR/CE FMR1 Reporter | N/A | 49576 |
T 1-877-777-1874; 512-681-5200 F 512-681-5202 E orders@asuragen.com
References
- Referenced in over 30 peer reviewed publications and used in over 200 laboratories, the AmplideX® PCR/CE FMR1 Reagents* are globally recognized as best-in-class for assessment of CGG repeats in the FMR1 gene.
- Key resources
- Videos
AmplideX® PCR/CE FMR1 Kit
AmplideX PCR/CE FMR1 Kit is an in vitro diagnostic (IVD) device for use in clinical laboratories for detection of the CGG repeats in the fragile X mental retardation (FMR1) gene. The device is intended to aid in the diagnosis of fragile X syndrome and fragile X associated disorders, e.g. tremor and ataxia syndrome (FX-TAS) and primary ovarian insufficiency (FXPOI), through determination of CGG repeat length up to 200 CGG and detection of alleles greater than 200 CGG. The kit provides a PCR-only approach based on Triplet Repeat Primed PCR (TP-PCR) design to reliably amplify and detect all alleles including Full Mutations.
Features & Benefits
AmplideX PCR/CE FMR1 Kit has created an easy-to-use, accessible, high performance method for laboratories to reliably analyze CGG repeats and detect interrupting AGG sequences in the FMR1 gene.
Reduced ComplexityEase-of-analysis of the FMR1 gene has been simplified through:
- Implementation of proprietary PCR solution for amplifying GC-rich regions
- Automation of result calling using AmplideX PCR/CE FMR1 Reporter
Optimized WorkflowValuable operator hands-on time has been significantly reduced through:
- Direct injection of PCR products (no PCR clean up) in to Capillary Electrophoresis platforms
- Decreased need for Southern blot analysis (up to 50 fold)
- End-to-end solution for FMR1 analysis including all necessary reagents and software
Quality PerformancePerforming FMR1 Analysis with Greater Sensitivity and Accuracy:
- Detection of all allele expansions, including low abundance full mutation size mosaics with up to at least 1300 CGG repeats
- Up to 875 fold more sensitive than Southern blot1
- Resolution of female homozygous and heterozygous samples and indication of interrupting AGG sequences
- Proven performance as indicated by more than 30 peer reviewed publications
Product Description
Analytical Characteristics of AmplideX PCR/CE FMR1 Kit:
- Proven clinical accuracy compared to Southern Blot (Table 1)
- Detects all alleles including low abundance full mutations (Figure 1)
- Accurately sizes all alleles up to 200 CGG repeats (Figure 2)
- Resolves female zygosity (Figure 3)
- Detects presence of AGG interruptions (Figure 4)
Table 1: Diagnostic Sensitivity of 100%; Diagnostic Specificity of 98.4% and Overall Accuracy of 99%*These 2 samples presented premutation alleles by both methods and low intensity full mutation alleles detected only by the AmplideX PCR/CE FMR1 Kit
Figure 1: Amplification of Asuragen’s Methylation and Sensitivity Control which has a 5% full mutation in a background of 95% Normal
Figure 2. Female Pre-mutation sample with accurate sizing of Normal (30 CGG) and Pre-mutation allele (56 CGG)
Figure 3: The difference in the “stutter” peak patterns of homozygous and heterozygous female provides a clear resolution of zygosity
Figure 4. Female Full Mutation sample with AGG interruptions as indicated by sudden decrease in peak heights of the “stutter” peak profile
Ordering
Product Name | Number of Reactions | Catalog Number |
---|---|---|
AmplideX PCR/CE FMR1 Control* | 24 UL | 49513 |
AmplideX mPCR FMR1 Control* | 24 UL | 49514 |
AmplideX PCR/CE FMR1 Reagents* | 100 | 49402 |
AmplideX PCR/CE FMR1 Kit | 100 | 76008 |
AmplideX mPCR FMR1 Kit* | 24 | 49442 |
AmplideX PCR/CE FMR1 Reporter* | N/A | 49576 |
T 1-877-777-1874; 512-681-5200 F 1-512-681-5202 E orders@asuragen.com
References
- Referenced in over 30 peer reviewed publications and used in over 200 laboratories, the AmplideX® PCR/CE FMR1 Reagents* are globally recognized as best-in-class for assessment of CGG repeats in the FMR1 gene.
- Key resources
- Videos
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我是这样进行预测的:在“SearchMode:”中选择“rRNAscanonly:”,在“sorce”中选择“mito/chloroplast”,粘贴序列,运行程序。得到了21个tRNA,而鱼中应该是22个tRNA,我觉得很奇怪,在“SearchMode:”和“rRNAscanonly:”中进行其它的设置,都不能得到22个tRNA.
我看的一篇文献“CompletemitochondrialDNAsequenceoftheJapanese
flyingfishCypselurushiraii”中也是说22个tRNA,但是我把文章提交的序列(Genbank号为AB182653.)拿到网上提交,只有21个tRNA,我觉得很奇怪。
不知道是怎么回事?请高手指点,那个软件好像是比较权威的。
中文名为信使RNA,是由DNA的一条链作为模板转录而来的、携带遗传信息的能指导蛋白质合成的一类单链核糖核酸。mRNA作为蛋白质合成的模板,决定肽链的排列顺序。
tRNA
中文名为转运RNA,是具有携带并转运氨基酸功能的一类小分子核糖核酸。
rRNA
中文名为核糖体RNA,是最多的一类RNA,也是3类RNA(tRNA,mRNA,rRNA)中相对分子质量最大的一类RNA,它与蛋白质结合而形成核糖体,其功能是作为mRNA的支架,使mRNA分子在其上展开,形成肽链的合成。rRNA占RNA总量的82%左右。rRNA单独存在时不执行其功能,它与多种蛋白质结合成核糖体,作为蛋白质生物合成的“装配机”。
n.[医] (对伤处等的) 针探,探查; [医] 探针,取样器; 探测仪;探头;
vt.探索,调查; 用探针(或探测器等)探查,探测;
vt.盘问; (用试探性袭击等)侦察(敌情) ; 用尖物刺穿(物件); 用力使向前推进;
The more they probed into his background, the more inflamed their suspicions would become
他们越调查他的背景,疑团就越多。
没法移动,也没有办法看信号强度啊?
siRNA (Small interfering RNA),是一种小RNA分子(~21-25核苷酸),由Dicer(RNAase Ⅲ家族中对双链RNA具有特异性的酶)加工而成。SiRNA是siRISC的主要成员,激发与之互补的目标mRNA的沉默。
PNA是一种人工合成的核酸的类似物,与核酸相似,它也是由携带ATGC四种碱基的四种单体首尾相连所构成的链状结构,从而保证这种物质能够像核酸一样靠碱基堆积力形成Watson-Crick碱基对,与DNA、RNA形成杂交分子PNA/DNA或PNA/RNA这就是PNA作为杂交探针的理论基础。所不同的是构成PNA骨架的单体不是通常的带负电的磷酸核糖而是N-(2-氨已基)-甘氨酸,单体之间靠酰胺键连接成长链。这种电中性的骨架结构赋予肽核酸探针优良的杂交特性。
首先PNA/DNA杂交分子的性质很稳定,从化学动力学角度看,PNA/DNA的结合常数有明显提高,一旦形成了双链,即使加入上百倍的寡核苷酸也不能使它解离。据报道,对于含有高度反向重复序列的超螺旋DNA,PNA/DNA杂交分子比DNA/DNA双链的结合常数提高了五万倍;从热力学角度看,PNA/DNA杂交分子的Tm也高于相同序列的DNA/DNA双链,通常,在100mmol/L的NaCl溶液中,PNA/DNA杂合子与相应的DNA/DNA双链相比,每个碱基对的Tm值要高出1℃。也就是说,15个碱基对的PNA/DNA平均Tm值约为70℃,比同样的DNA/DNA要高出15℃。从而使检测过程中的灵敏度大为提高。此外,中性骨架还使PNA与DNA的杂交摆脱了对杂交液盐度的依赖,这一重要性质不仅意味着杂交过程中省去了提高盐浓度的操作,更有意义的是,只要保证杂交体系足够低的离子强度,就能抑制非特异性的杂交和靶DNA序列的自身复性对灵敏度的损害。
不但如此,这种高度的灵敏度并没有像其他种类的探针一样引起特异性的下降,相反它对于碱基错配的容纳力更小。在16bp的PNA/DNA杂合子中,一个碱基的错配会导致Tm值下降15℃,而对于同样的DNA双链,Tm值只下降11℃
原文地址:http://tahepna.com/file_news_read.asp?id=59&class=%D1%D0%BE%BF%BD%F8%D5%B9&fo=4&flag=
大家好,近期在用lipo2000做悬浮细胞HL-60的siRNA转染,siRNA是由公司合成的三个siRNA,但是敲除效率很低,并且每次qPCR后做出来的结果都不一样:有时是第一个siRNA敲除作用好点,但有时是第二个或第三个siRNA效果好。由于结果不稳,并且细胞不好转我就将细胞换成了A549,但是换成了贴壁细胞后发现每次转染后进行qPCR检测时结果还是不稳,另外,我还设置了不同siRNA的组合,有时结果显示siRNA1+siRNA3效果好,但有时显示siRNA1+2+3效果好。光做这个siRNA转染已经几个月了,到现在都没有确定具体哪个片段起作用或效果最好,也没有一个稳定的趋势。请问造成这种敲除效率不稳定的原因到底是为什么?急切等待回复,谢谢!
tRNA(转运RNA)可以转运氨基酸。
mRNA(信使RNA)是由细胞核内的DNA转录来的,相当于蛋白质的设计图纸。
翻译的过程:核糖体附着在mRNA上,然后tRNA搬运氨基酸,运往到核糖体上,从而拼合成蛋白质。
编码线虫的氨基酰tRNA合成酶是否和编码大鼠的氨基酰tRNA合成酶的基因相同?
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