EasyYACPreparationMethod
YAC TRANSFORMATION OF C. ELEGANS USING TOTAL YEAST GENOMIC DNA[This method is described in The Worm Breeder's Gazette (1997) A. Davies and J. Shaw, 15(1)]1. PREPARING YAC (+ Yeast) DNA BY CsCl PURIFICATION.(Adapted from Stiles (1983) Methods in Enzymology 101: 296-297)Grow 3ml culture of yeast strain carrying your YAC of interest in selective media (e.g. CM URA- TRP-) @ 30 C for 1-2 overnights.Add culture to 50ml......阅读全文
Easy－YAC－Preparation－Method
YAC TRANSFORMATION OF C. ELEGANS USING TOTAL YEAST GENOMIC DNA[This method is described in The Worm Breeder's Gazette (1997) A. Davies and J. Shaw
酵母人工染色体
· Easy YAC Preparation Method (Andrew Davies,Shaw lab)· Screening YAC libraries (Donis Keller Lab)This is a method for screening YAC l
Column－Method－for－Lambda－Phage－DNA－Preparation
Purpose:Mini-prep method for lambda phage DNA purification from lysates.Time required:4 hours once the lysate is in handSpecial supplies required:BioR
Method:－Preparation－of－Lymphocyte－Cell－Pellet－for－Storage
Method: Preparation of Lymphocyte Cell Pellet for StorageJune 10, 1990Rosalie VeilePurpose:Following propagation to 1 X 108 cells, lymphoblastoid cell
Method:－Preparation－of－Lymphoblastoid－Cell－Lines－for－Long－Term－Storage
Method: Preparation of Lymphoblastoid Cell Lines for Long Term StorageMay 30, 1990Rosalie VeilePurpose:To store cell lines in a form that will insure
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells2
Flow Cytometric AnalysisPrior to flow cytometric analysis, the vital dye Hoechst 33342 (Sigma-Aldrich, St. Louis, MO, USA) was added to cell suspensio
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells3
Moreover, when the method was applied to the analysis of the cellular composition of immature testes, the results were also in agreement with previous
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
Optimized－Method－for－the－Preparation－of－Rodent－Testicular－Cells1
Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the c
A－practical－method－for－the－preparation－of－total－DNA－from－filamentous－fungi
Most methods of DNA preparation from fungi are time-consuming due to the need to first make protoplasts, expensive for chemicals such as cesium chlori
YAC实验——YAC文库筛选
实验步骤1. YAC中心实验室用于筛选文库的方法进展很快。2. 将一块或多块微量滴定板微孔中的YAC克隆转印到尼龙膜上，用单拷贝探针进行杂交，就可以筛选YAC文库了。3. 然而，由 于杂交实验的信噪比较低以及制备所有的转印尼龙膜所需的花费巨大，绝大多数实验室已采用PCR方法来筛选文库。4.
YAC实验——YAC文库构建
YAC带有天然染色体所有的功能元件，包括一个着丝粒，一个DNA复制起点，两个端粒。YAC能够容纳长达几百kb的外源DNA，这是质粒和黏粒办不到的。大片段的插入更有可能包含完整的基因，在染色体步移中每次允许更大的步移距离，同时能够减少完整基因组文库所需的克隆数目。实验步骤1. 尽管当插入片段大于10
YAC实验
1. 尽管当插入片段大于100 kb 时，YAC克隆是可供选择的基因组DNA大片段克隆方法之一，但该系统的一些特性使之很难被迅速采纳为常规克隆手段。2. 在复杂的酿酒酵母菌的基因组中，通常所用的YAC只是一个单拷贝。3. 因此在YAC文库中鉴定一个克隆，其信噪比较之在λ噬菌体载体或粘粒载体文库
YAC实验
YAC文库构建 YAC文库筛选 实验步骤 1. 尽管当插入片段大于100 kb 时，YAC克隆是
Southen杂交
Southern杂交One important thing for transfer:the weight of the object resting on top of the blotting apparatus should not exceed the weight equal to a 5
组织学——显微解剖
Laser Capture Microdissection (LCM)Introduction to LCM (BJMU) Preparation, LCM and RNA/DNA extraction of Frozen Tissue Sections (NIH Laser Capture M
YAC克隆的分析实验——制备YAC－DNA进行Southern印迹分析
实验材料酵母菌试剂、试剂盒AHCSCESCEMTris·ClEDTATE乙酸甲RNase A异丙醇NaCl仪器、耗材培养箱离心管转子实验步骤1. 接种一个红色的含YAC克隆的酵母菌落于盛有20 ml AHC培养基的250 ml 三角烧瓶中，在旋转摇床上30℃ 250 r/min 振荡培养24 h。
组织学——组织制备
· Histological techniques (William H. Heidcamp)Very detailed guide to histological techniques, like fixation, dehydration, embedment and subs
Quantification－made－easy
With the aid of spectroscopy, the quantitative analysis of nucleic acids and proteins has established itself as a routine method in many laboratories.
YAC克隆的分析实验
实验材料 酵母菌试剂、试剂盒 AHCYPD甘油仪器、耗材 培养箱冷冻管实验步骤 1. 用接种棒将含重组载体pYAC的AB1380菌株划线接种在AHC平板上，倒扣平板于30℃培养，直至长出1~3 mm 大小的红色菌落为止。2. 短期保存：用Parafilm膜将平板周围封起来，于4℃可保存4~6周。
YAC克隆的分析实验
基本方案 制备YAC DNA进行Southern印迹分析 实验材料 酵母菌
Vacuum/Spin－Protocol－for－Tissue－DNA－Extraction
实验概要The E.Z.N.A.® Tissue DNA Kit provides a rapid and easy method for the isolation of genomic DNA for consistent PCR and Southern analysis. Up to 3
碳水化合物分析
Carbohydrate Assay (Hancock Laboratory) (Accessible only by IE)This protocol is used to determine the relative amounts of LPS CHO present in a given s
细胞遗传学——原位杂交(ISH)
In Situ Hybridization· In Situ Hybridization (jsmith1@po-box.mcgill.ca)In situ hybridization, as the name suggests, is a method of localizing,
Preparation－of－tubulin
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
Template－Preparation
Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template. ABI recommends a minialkaline-lysis/PEG preci
Platelet－Preparation
OUTLINEIn order to avoid platelet activation all manipulations must be performed as quickly and as acurate as possible.Work on ice if possible! This p
Liposome－Preparation
Liposome PreparationOBJECTIVE:Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation m
CAM－preparation
8 eggs per day, day 7- day 13 cut CAM, wash in precooled PBS,in 10 ml WASH 1 (PBS, 5 mM EDTA, COMPLETE) on icecut in pieces in petri dish on icecentri
SMEAR－PREPARATION
The preparation of a smear is required for many laboratory procedures, including the Gram-stain. The purpose of making a smear is to fix the bacteria