LargeScalePlasmid,Cosmid,BAC,PAC,andFosmidDNAIsolation
DNA Isolation by a Cleared Lysate Method Followed by Double Acetate Precipitation Version 3b - updated September 26, 1999The Most Recent Roe Lab Implementation (by Feng Chen, Hau-Qin Pan and Fu Ying) Adapted from the Genome Sequencing Center, Washington Univ. ProtocolA smear of colonies of cosmid/BAC/PAC were picked and transferred into a 12X75 mm Falcon tube containing 3 ml of LB medium (10 g Bacto-Tryptone, 5 g Bac......阅读全文
用杂交的方法筛选大片段插入的文库实验
实验步骤由于原代的转化子只能产生数目有限的滤膜，因而随机铺板不适合滤膜的制备以及 YAC、BAC 和 PAC 文库的保存，而且由于克隆增殖率存在差别，因而无论是哪一种扩增形式都会将偏差带人文库中去。因此，原代的转化子应加入微孔板中，放置于-8℃ 保存。这使得制造分配用的文库拷贝变得简单易行，而且使组
用杂交的方法筛选大片段插入的文库实验
由于原代的转化子只能产生数目有限的滤膜，因而随机铺板不适合滤膜的制备以及 YAC、BAC 和 PAC 文库的保存，而且由于克隆增殖率存在差别，因而无论是哪一种扩增形式都会将偏差带人文库中去。因此，原代的转化子应加入微孔板中，放置于-8℃ 保存。这使得制造分配用的文库拷贝变得简单易行，而且使组
High－Molecular－Weight－Yeast－Liquid－DNA－Preparation
Purpose:To isolate intact, high molecular weight DNA from yeast cells for subcloning and rare cutting restriction enzyme analysis. One can expect a yi
Purifying－Large－E.－coli－Restriction－Fragments－from－PulsedField－Gels
DNA PreparationE. coli chromosomal DNA is prepared following the method of Heath et al. ( J. Bacteriol., 174, 1992). Cells are embedded in agarose, th
Isolation,－Culture,－Characterization－of－Cortical－and－Hippocampal－Neurons
实验概要The ability to culture primary neurons under serum-free conditions facilitates tighter control of neuronal studies. Some serum-free media and s
Isolation－of－human－primary－airway－smooth－muscle
1. Primary cultures of human primary airway smooth muscle (ASM) cells were prepared from explants of airway smooth muscle. 2. The human trachea tiss
Isolation－of－human－colonic－epithelial－cells－(EC)
1.Human colonic epithelial cells (EC) were isolated from freshly resected colonic surgical specimens obtained. 2.All diagnoses were confirmed by clini
Isolation－of－osteoblasts－from－human－bone.
Bone samples were cleaned of adherent soft tissue and osteoblasts isolated by two methods.a. The bone was cut into small pieces (2 mm × 2 mm), washed
Isolation－of－retinal－pigment－epithelial－cell（一）
The pigmented layer of retina or retinal pigment epithelium (RPE) is the pigmented cell layer just outside the neurosensory retina that nourishes
Isolation－and－Culture－of－Human－Liver－Stem－Cells
Materials and Methods1. Human hepatocytes were isolated from fresh surgical specimens of patients undergoing hepatectomies. Healthy liver tissue (
PriCells:－Isolation－of－endothelial－progenitor－cells－(EPCs)
PriCells: Isolation of endothelial progenitor cells (EPCs) 1. Twenty-four mL venous blood was collected at each time point into Vacutainer CPT Mono
Isolation－and－cultivation－of－endothelial－progenitor－cells－(EPCs)
Circulating bone marrow (BM)–derived endothelial progenitor cells (EPCs) are recruited to the site of tissue regeneration and substantially contri
CD133－isolation,－expansion,－and－differentiation
1. Renal progenitor cells were obtained from the normal portion of cortex obtained from surgically removed kidneys. 2. After dissection and passage th
Mouse－B－cell－isolation－with－magnetic－beads
实验概要Isolate B cells from total mouse cells实验步骤 Praparation:1. Enough MACS; put RPMI into water bath.2. Put some MACS in 15 ml tube or 1.5
Isolation－of－peripheral－blood－mononuclear－cells－(PBMC)
1. Acid-citrate dextrose-treated blood (450 ml) was obtained from donors. 2. PBMC were isolated by centrifugation over lymphocyte separation medium. 3
Isolation－of－human－prostatic－smooth－muscle－cells
Human prostate tissues1.Human hyperplastic prostates were obtained during surgery from four men through transurethral resection of the prostate. 2.All
Primary－type－II－pneumonocyte－Isolation－and－culture
Pulmonary alveolar type II cells carry out highly specialized functions that include the synthesis, secretion, and reutilization of surfactant, a
Isolation－of－human－uterine－smooth－muscle－cells
1. Approval to utilize human tissue was given by Hospital's Committee on Research Involving Human Subjects. 2. Human myometrial and leiomyoma tiss
Isolation－and－growth－of－pulmonary－artery－adventitial－fibroblasts
1. Adventitia from the main pulmonary artery was harvested neonatal calves. 2. Tissue was collected, carefully dissected free of blood vessels and f
Isolation－and－characterization－of－rat－glomerular－endothelial－cells
Glomerular endothelial cells (GECs) from the kidney are in close juxtaposition to other cell types, such as mesangial cells, and may respond to as
Isolation－of－human－corneal－endothelial－cells－(HCECs)
Isolation of HCECs1. The corneoscleral tissues were rinsed three times with primary cell culture system containing 50 mg/mL gentamicin and 1.25 mg
Isolation－of－retinal－pigment－epithelial－cell（二）
15. This medium is replaced after 1 day with 5% serum-containing RPE medium, and subsequent changes were made every 2 to 3 days.16. After 3 to 4 weeks
PAC1基因编码的功能和结构描述
该基因编码的蛋白是一种GTP酶，属于小GTP结合蛋白的ras超家族。这个超家族的成员似乎调节着各种各样的细胞事件，包括控制细胞生长、细胞骨架重组和蛋白激酶的激活。两个编码不同亚型的转录变体已经被发现。The protein encoded by this gene is a GTPase which
PAC2基因编码的功能和结构描述
该基因编码小鸟苷三磷酸（gtp）代谢蛋白ras超家族的一个成员。编码的蛋白质定位于质膜，在质膜上调节各种过程，如分泌、吞噬和细胞极化。这种蛋白质的活性也与活性氧的产生有关。该基因突变与中性粒细胞免疫缺陷综合征有关。这个基因在第6号染色体上有一个假基因。This gene encodes a memb
PAC3基因的结构特点和主要作用
该基因编码的蛋白是一种GTPase，属于小GTP结合蛋白RAS超家族这个超家族的成员似乎调节了一系列不同的细胞事件，包括控制细胞生长、细胞骨架重组和激活蛋白激酶选择性剪接导致多个转录变体
PAC3基因编码的功能和结构描述
该基因编码的蛋白是一种GTPase，属于小GTP结合蛋白RAS超家族这个超家族的成员似乎调节了一系列不同的细胞事件，包括控制细胞生长、细胞骨架重组和激活蛋白激酶选择性剪接导致多个转录变体。The protein encoded by this gene is a GTPase which belon
PAC3基因的结构特点和主要作用
该基因编码的蛋白是一种GTPase，属于小GTP结合蛋白RAS超家族这个超家族的成员似乎调节了一系列不同的细胞事件，包括控制细胞生长、细胞骨架重组和激活蛋白激酶选择性剪接导致多个转录变体
Sauer:Lysing－E.－coli－with－Lysozymes
Getting The Most Out Of Your BugsNative lysis is a staple protocol in practically every biochemistry lab, yet there is significant variability in the
pEZ™－BAC载体的基本信息和质粒图谱
pEZ™ BAC载体载体基本信息载体名称pEZ™ BAC载体抗性Chloramphenicol载体长度7260 bp载体类型Basic Cloning Vectors载体来源Lucigen拷贝数Low copy numberpEZ™ BAC载体质粒图谱
细菌的核酸抽提
DNA Extraction· DNA Extraction from Bacteria (Julie B. Wolf,UMBC)Phenol/chloroform method· DNA Extraction From Bacteria (Triton Method