摘要: 10X TBE, per literSequencing dye-----------------------------------108g Tris base80% formamide55g boric acid10mM NaOH40ml 0.5M EDTA1mM EDTA0.1% xylene cyanol10% APS0.1% bromophenol blue--------------[阅读全文]

摘要:MediaNeed LB plates (9 cm or 14 cm)to plate the library and do secondary screens.I typically screen about 500,000-1 million plaques for a primary cDNA screen.This fits on 11-22 14 cm plates.I do the s[阅读全文]

摘要:Plate a bacteriophage library at about 30,000-50,000 pfus per 150mm Petri dish by adding an appropriate amount of bacteriophage to 500µl MgSO4-stored E.coli.Dilute bacteria,if needed(ex.VCS237 t[阅读全文]

摘要:S1 Nuclease Protection Assay End-label oligo (20-25 mer)4 µl 5X T4 Kinase Buffer5 µl [gamma-32P] ATP (7000 Ci/mmol)10 µl water + oligo (200 ng)1 µl T4 Kinase1. 37 deg C for 1 h[阅读全文]

摘要:1. Run gel (0.8 -1.0% agarose is best). For yeast chromosomal Southerns, digest 20 μg DNA. Use 50 ml minigel for most purposes. Photograph gel, but minimize exposure to UV light.2. Depurination: Pl[阅读全文]

摘要:Digest DNA in 96-well plate To each well add:4μl 10Xbuffer4μl Enzyme0.4μl Spermidine(0.4M)31.6μl H2 O37℃ 19h, then add 4μl loading dye to each well. Load into 400ml 1% agarose gel immed[阅读全文]

摘要:Southern Hybridization Protocols Two protocols are given here.The first one is used for BAC library screening on filters, although is also suitable for Transfer hybridizations. PREPARATION OF HYBRIDIZ[阅读全文]

摘要:Hello,I have been trying over the last few months to introduce a single point mutation in a looped portion of an RNA, using the dut-ung method of mutagenesis.So far, I have had little success.I have a[阅读全文]

摘要:Contributed by Dr. A. GratchevSingle Nucleotide Primer Extension is a powerful method which can be used for the precise analysis of methylation in a certain position. The procedure is shown on the fig[阅读全文]

摘要:Hi can anyone answer thisI am having problems with my southern hybridization.I am wondering if the DNA transfers to my membrane.Is it possible that the DNA can wash into the solutions when denaturing [阅读全文]

摘要:10X TBE, per literSequencing dye-----------------------------------108g Tris base80% formamide55g boric acid10mM NaOH40ml 0.5M EDTA1mM EDTA0.1% xylene cyanol10% APS0.1% bromophenol blue---------------[阅读全文]

摘要:Hey everybody,I am working on a side-directed mutagenesis with the Stratagene℃ quickchange mutagenesis xl-kit. Here is my problem: My plasmid is 17kb big. Until now I have had no colonies on my plate.[阅读全文]

摘要:Paula Marquardt & Craig EchtPublished in:Echt CS, May-Marquardt P, Hseih M, Zahorchak R. 1996. Characterization of microsatellite markers in eastern white pine. Genome 39:1102-1108 .comments can be di[阅读全文]

摘要:This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure with a variety of Populus species, as well as tobacco and[阅读全文]

摘要:Materials:Silica Suspension:add 2 g of silica to 15 ml of H2Owash 3x by centifugation at 2000 x g for 2 minestimate vol of silica and resuspend in 2 vol H2OSilica Wash Solution:50 mM NaCl, 10 mM Tris [阅读全文]

摘要:一 酶切和电泳在200 μl 微量离心管中加入：25 μl DNA样品(约10μg)，3μl 限制性内切酶（MBI，10 U/ μl）5 μl 相应的10×buffer，补水到50μl。然后加一滴矿物油覆盖, 稍微离心后放于37℃水浴8-12小[阅读全文]

摘要:This kit contains materials for six groups to perform Southern transfer and hybridization analysis using the included lambda DNA samples and biotinylated probe. The intellectual objective of the exper[阅读全文]

摘要: PurposeDNA extraction without phenol extraction and centrifugation.Procedure1. Lysis: the lysis buffer (usually 0.5ml) is added to the tissue or cell ( for a 75cm2 flask, add 5ml directly to the cell[阅读全文]

摘要:本方法由Dellaporta,Wood和Hicks(1983)的方法修改而成。其基本原理是研磨的组织细胞用热的SDS裂解后，加入高浓度的KAc，0℃放置以除去蛋白和多糖类杂质，最后用乙醇或异丙醇沉淀。一 材料、试剂和仪器1 材料 新鲜的组织材料[阅读全文]

摘要:Has anyone any experience of using a semi dry blotter for Southern blotting. At present I am tranfering overnight using a weight, I think there is a blotter that is suitable for nucleic acids and can [阅读全文]