扩增反应的条件在50μl的反应体积中含有50mmol/L KC1,10mmol/L Tris-Cl,(pH8.5),1.5mmol/L MgCl2,200μmol/L dNTP,10pmol引物,1U的Taq DNA聚合酶(Clontech公司产品)。在PE9600型DNA热循环仪(Perkin-Elmer公x司)上按下列条件反应25个周期94℃30sec;55℃30sec;72℃2min。在RT-PCR时同时设β-actin为阳性...
实施例4重组人丝氨酸-tRNA合成酶56.65的体外表达、分离和纯化根据SEQ ID NO1和 图1所示的编码区序列,设计出一对特异性扩增引物,序列如下Primer35’-CATGCTAGCATGCTCTTATGTGATAAAGATGAG-3’(Seq ID No5)Primer45’-CCCGAATTCCTATATCCTCTTCAGTATGGTGAT-3’(Seq ID No6)此两段引物的5’端分别含有NheI和EcoRI酶...
扩增反应的条件在50μl的反应体积中含有50mmol/L KCl,10mmol/LTris-Cl,(pH8.5),1.5mmol/L MgCl2,200μmol/L dNTP,10pmol引物,1U的Taq DNA聚合酶(Clontech公司产品)。在PE9600型DNA热循环仪(Perkin-Elmer公司)上按下列条件反应25个周期94℃ 30sec;55℃ 30sec;72℃ 2min。在RT-PCR时同时设β-actin为阳性...
Sec-TCA tdbD00011827 AS-NR-001-1-43 Ser-ACT tRNA-Ser-ACT-1-1 AS-NR-001-1-44 Ser-AGA tdbD00009824 AS-NR-001-1-45 Ser-CGA tdbD00009832 AS-NR-001-1-46 Ser-GCT tdbD00009833/tdbD00009834/tdbD00009835/tdbD00009836/tdbD00009837 AS-NR-001-1-47 Ser-GGA nmt-tRNA-Ser-...
Filters were washed with 2 mL of 5% TCA and 3 mL of 95% EtOH and dried. Liquid scintillation counting was used to quantify the amount of t6A-modified tRNA using the standard curve to convert CPM to pmol of tRNA (1 pmol of tRNA = 97.8 CPM). Fluorescence anisotropy measurements ...
在genbank中使用索取号HQ324232搜索并下载到该物种Nanorana pleskei的线粒体基因组保存为文件Nanorana pleskei.fasta 登录tRNAscan-SE服务器 http://lowelab.ucsc.edu/tRNAscan-SE/ 在搜索模型（Search Mode）中选择“tRNAscan-> Cove”，数据来源（Source）中选择“Mito/Chloroplast”在提交序列文件选项（submit a ...
sieinnmgeitlrraenrciWrtyu.1i)TtmihneeSnottbapvtirilooiaucesspsss.e,TqnhuaeemntrecnleyWstirm1naiRnladcroittnrynvoWefrtt2reondfWtSot2atartninlidWatsnbpry.Rhp(ooFlidingtt. hm2eCus)taaimntideoinacnaattteictshotedhotehndirubdpulatincshatitocewotdrtnohRne position from triplet "TCG" to "TCA"...
Supernatant represents selenium present in the form of low molecular weight selenocompounds after removal of protein, and the trichloroacetic acid (TCA) fraction represents selenium present in the form of proteins. SampleGenotypeSelenium levelsa ppm Total liver extract Trspfl/fl 1.14 Alb-Cre+/- 0.99...
在PE9600型DNA热循环仪(Perkin-Elmer公司)上按下列条件反应25个周期94℃ 30sec;55℃ 30sec;72℃ 2min。在RT-PCR时同时设β-actin为阳性对照和模板空白为阴性对照。扩增产物用QIAGEN公司的试剂盒纯化,用TA克隆试剂盒连接到pCR载体上(Invitrogen公司产品)。DNA序列分析结果表明PCR产物的DNA序列与SEQ ID NO1所示的1...
PrimeSTAR GXL DNA polymerase (TAKARA) was used in the long PCRs with the cycling conditions: 98 °C for 10 sec, 68 °C for 2 to 10 min (depends on the length of regions between rrnL and cox1). The reaction mixture contained 0.5 μ l GXL DNA Polymerase, 5 μ...