今天窝牛号就给我们广大朋友来聊聊primera,以下关于的观点希望能帮助到您找到想要的百科。

最佳回答搜集了几个植物RT-PCR的内标,与各位共享

1． 物种：拟南芥（arabidopsis）或maize

2． 基因名称：18S rRNA

3． 模板：cDNA

4． PCR类型及目的：RT-PCR

5． Forward primer (5’-3’): CCATAAACGATGCCGGA

Reverse primer (5’-3’): CACCACCCATAGAATCAAGA

Probe: 无

6． 产物长度（bp）：350

7． 引物浓度：50 pmol/ul

8． 退火温度：54.1

9． 所用仪器：PE-9600

10． 提交者ID：yuxing955

11． 提交者辩闷联系方式：xing-yu955@163.com

12． 参考文献原文：本人亲自验证，还有本实验室其他人亲自验证。

1． 物种： maize

2． 基因名称：actin

3． 模板：cDNA

4． PCR类型及目的：RT-PCR

5． Forward primer (5’-3’): AAATGACGCAGATTATGTTTGA

Reverse primer (5’-3’): GCTCGTAGTGAGGGAGTACC

Probe: 无

6． 产物长度（bp）：

7． 引物浓度：50 pmol/ul

8． 退火温度：54.1

9． 所用仪器：PE-9600

10． 提交者ID：yuxing955

11． 提交者联携档弯系方式：xing-yu955@163.com

12． 参考文献原文：本人亲自验证，还有本实验室其他人蠢判亲自验证

Semiquantitative RT-PCR

Reverse transcriptions were carried out using the Super Script II RT and the adapter primer of the 3' RACE System. For amplification of ROT4 cDNA, ROT4-F and ROT4-R were used. As a control, the following oligonucleotides were used to detect the constitutively expressed ACTIN2 (ACT2) gene (An et al., 1996): ACT2-F, 5'-GAAATCACAGCACTTGCACC-3'; ACT2-R, 5'-AAGCCTTTGATCTTGAGAGC-3'. The condition for amplification by RT-PCR was one cycle at 50°C for 15 min and 94°C for 2 min, then 27 cycles for ROT4 cDNA amplification or 33 cycles for ACT2 cDNA amplification at 94°C for 15 sec, 57°C for 30 sec, and 72°C for 60 sec.

可以

The Arabidopsis AtACT8 (actin 8) gene was used as a positive internal control (An et al. 1996). The PCR primers used to detect AtACT8 mRNA were 5'-ATG AAG ATT AAG GTC GTG GC-3' and 5'-TCC GAG TTT GAA GAG GCT AC-3' (Aida et al. 1997). These products were fractionated by electrophoresis on a 3% agarose gel and stained with ethidium bromide, and scanned using a fluorescent image analyser.

。

UBQ, 5_-GATCTTTGCCGGAAAACAATTGGAGGATGGT-3_

and 5_-CGACTTGTCATTAGAAAGAAAGAGATAACAGG-3_.

Quantitative PCR Analysis of GSL mRNA

Total RNA (5 μg) was used in cDNA reactions using the Superscript II cDNA synthesis kit (Invitrogen). The cDNA was diluted 2.5-fold, and 1 μL was taken for quantitative real-time PCR in 20-μL reaction volumes using 10 μL of 2× Quanti-Tect PCR master mix (Qiagen, Valencia, CA), 0.3 μM gene-specific primers, and 0.6 μL of a 100-fold dilution of SYBR Green I dye (Applied Biosystems, Foster City, CA). PCR cycling and fluorescence measurements were performed with a Rotorgene 2000 Real-Time Cycler RG2072 (Corbett, Sydney, Australia), and data were normalized against glyceraldehyde phosphate dehydrogenase (GAPDH), actin (Actin1), and cyclophilin (Cyclo) mRNA levels (Vandesompele et al., 2002 ). Primers used in the quantitative PCR were as follows: for AtGAPDH (At3g26650), 5′-TGGTTGATCTCGTTGTGCAGGTCTC-3′/5′-GTCAGCCAAGTCAACAACTCTCTG-3′; for AtActin1 (At2g37620), 5′-TGCGACAATGGAACTGGAATG-3′/5′-GGATAGCATGTGGAAGTGCATAC-3′; for AtCyclo (At2g36130), 5′-TGGCGAACGCTGGTCCTAATACA-3′/5′-CAAAAACTCCTCTGCCCCAATCAA-3′; for AtGSL5 (At4g03550), 5′-CTGGAATGCTGTTGTCTCTGTTG-3′/5′-TCGCCTTTTGATTTCTTCCCAGT-3′; for AtGSL6 (At1g05570), 5′-GAAGGGTTTGGGCGTTGGAAG-3′/5′-CAATGAGAAGCATTCCCCATCCAGTT-3′; and for AtGSL11 (At3g59100), 5′-TTTAGGGGTTTGGGACTCGGTGAAA-3′/5′-TGTCTTTCCGACCAGCGAGAATCA-3′.

最佳回答编译器不会迟明有问题, 应该STL的问题

在我VC9.0上编译没什么问题.

执行svec.assign(slist.begin() , slist.end())的核心语句是

for (; _First != _Last; ++_Dest, ++_First)

_Al.construct(_Dest, *_First); // first等于list::iterator类型, 解开就等于char* p = "mary"

_Al.construct(_Dest, *_First);展码搜告开等于

::new (_Vptr) _T1(_Val); //_Val = char*类型 = "mary", _T1 = string类型

所以string( "mary" )完全是合法调用.

而你的编译器提示list::iterator无法转换成string*类型. 这实在让人费解,

我不相信你的STL会有类漏绝似string* p = list::iterator的间接操作.

是不是你STL文件被无意改过了.

比如删掉了*解引用操作符.

因为我删掉了*后, 我的编译器显式错误如下:

>d:\vc++\vc\include\memory(129) : error C2664: “std::allocator<_Ty>::construct”: 不能将参数 2 从“std::list<_Ty>::_Iterator<_Secure_validation>”转换为“const std::string &”

吓尿了.

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