摘要:Kitto Lab, The University of Texas at Austin http://research.cm.utexas.edu/bkitto/Kittolabpage/Protocols/Microbiology/electroporation.html This procedure prepares glycerol stock cultures of bacteria f[阅读全文]

摘要:The Preuss Lab,The Division of Biological Sciences,The University of Chicago http://preuss.bsd.uchicago.edu/protocols/topo.html [阅读全文]

摘要:Transformation DNA fragments (or plasmid DNA) into competent E. coli * Caution: Use aerosol protecting tips if selection of transformed cells is not based on X-gal strategy.1.Remove competent cells (E[阅读全文]

摘要:Donis-Keller Lab Manual, Department of Genetics in Washington University School of Medicinehttp://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid08.htmlPurpose: To utilize competent E. coli bacteria to rep[阅读全文]

摘要:Preparation of electroporation cells1. Prepare an overnight of NM522 in minimal medium.2. Inoculate 1L LB with 10ml (1/100th vol) of the overnight and grow to A600 = 0.5- 1.0.3. Pellet cells 5krpm, 15[阅读全文]

摘要:NOTE: THIS IS FINE FOR SOUTHERNS, BUT NOT FOR SCREENING BY PCR. (from Ruixia, 7/99, from protocol by Stef Oehen 7/94). 1. Put 1 cm tail in 1.5 ml microcentrifuge tube.2. Add 500µl Tail Buffer an[阅读全文]

摘要:About 1/2 to 1 cm of tail should be cut from properly marked mice (using toe or ear clipping, etc), about the age of 2-3 weeks. Place these tails into marked 1.5 ml Eppindorf tubes for processing; the[阅读全文]

摘要:1. 细菌离心后，加入溶液I蜗旋振荡时，发现菌体呈絮状不易混匀，或成细砂样。—— 很可能是细菌发生溶菌，可减少培养时间、降低培养温度试试看，通常降低培养温度 会使细菌生长更加稳定；同时也可以试试平板培养 ，[阅读全文]

摘要:问题：转化后无克隆菌产生 可能的原因：转化过程有问题或感受态细胞失活 建议：可通过转化pUC18/19等未切割的可用于抗性筛选的质粒，检测感受态细胞的转化效率和转化操作是否正确 问题：插入对照DNA片段的阳性率低 可能[阅读全文]

摘要:1.Grow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube). 2.Resuspend pellet with 300μl STET buffer (900μl). After resuspending add 30μl RNase/lysozym[阅读全文]

摘要:Transgenic Mouse and Gene Targeting FacilityTg 008 Southern BlottingMaterials and Equipment Hybaid hybridization oven (ISC BioExpress,H-9250)Vacuum oven (VWR,52201-218)Ludlum Geiger CounterRadiation S[阅读全文]

摘要:BUFFERS:1) Prehybridization/hybridization buffer1% SDS6X SSC10 % dextran sulfate100mg salmon sperm DNA/ml (added with the radioactive probe)Note: Store hybridization buffer at 4 degrees and warm to 65[阅读全文]

摘要:TRIzol试剂适用于从细胞和组织中快速分离RNA。TRIzol试剂有多组分分离作用，与其他方法如硫氰酸胍/酚法、酚/SDS法、盐酸胍法、硫氰酸胍法等相比，最大特点是可同时分离一个样品的RNA\DNA\蛋白质.TRIzol使样品匀浆化，细胞裂[阅读全文]

摘要:A claymation of the assembly of the spliceosome, and subsequent mRNA splicing." The Spliceosome Dance（剪切子的模拟动画） "[阅读全文]

摘要:本制品是一种快速方便的总RNA提取试剂，可以从动物组织、植物材料、各种微生物、培养细胞等中提取总RNA。样品在RNAiso Reagent中能够充分被裂解，在加入氯仿离心后，溶液会形成上清层、中间层和有机层（下层)，RNA分布在上清层[阅读全文]

摘要:The Easiest Route to Guaranteed SilencingIncreasing numbers of researchers are using small interfering RNAs (siRNAs) to reduce the expression of specific mammalian genes. Applications of siRNA induced[阅读全文]

摘要:The Easiest Route to Guaranteed SilencingIncreasing numbers of researchers are using small interfering RNAs (siRNAs) to reduce the expression of specific mammalian genes. Applications of siRNA induced[阅读全文]

摘要:Transcription of cRNA probeReagents/SolutionsDNA (PCR or plasmid) with bacteriophage RNA polymerase promoter in suitable orientation (see note 1 ) ~1µg/µl in TE buffer 10mM MgCl2 5x Transc[阅读全文]

摘要:Here is our T7 siRNA protocol. The main idea is to design primers that begin with GG so thattranscription by T7 polymerase is efficient.I) FIRST design oligos:Using sense coding sequence of any gene..[阅读全文]

摘要:TRIzol method for RNA isolation-To tissue or cells, add 200- 1 ml TRIzol (GIBCO BRL)1.homogenize for 60 sec (vortex or polytron)2.add 40ul-200 ml chloroform (1/5 volume of Trizol)3.mix by vortexing fo[阅读全文]