CHOCentrosomePrep
CHO Centrosome Prep:Arshad Desai4/94Cells:We grow our CHOs with MEM[[alpha]] (without nucleosides) + 10% Bovine Calf Serum and penn/strep/glutamine. For a prep it is best to grow twenty large plates (150 mM) and the cells should be grown to overconfluence - till they start piling up on each other (you will need to feed them frequently to keep them happy)Protocol Rationale:The protocol is identical to Tim's publis......閱讀全文
CHO－Centrosome－Prep
CHO Centrosome Prep:Arshad Desai4/94Cells:We grow our CHOs with MEM[[alpha]] (without nucleosides) + 10% Bovine Calf Serum and penn/strep/glutamine. F
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through
細胞組分和細胞器——染色體
Chromosomal DNA Prep : cultured cells/tissue samples?(Mike A Dyer)This protocol was developed for cultured cells but should be appropriate for dissoci
TRIzol－Prep
Procedure1.? Homogenize cells (10 million) or tissue (50-100 mg) in 1 mL?TRIzol Reagent?(e.g. scrape and pass through 30G needle, dounce homogenize an
Studier－Lysate－Prep
Summary How to make a lysate from a plaque preparation. We also use this protocol for preparation of a quick stock from previously made lysate prep.
Yeast－DNA－Prep
Protocolgrow up yeast culture to appropriate density (near saturation)spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatantresus
Competent－agro－prep－for－electroporation
day 1 1. Start 75 mL overnight cultures of agro (strain GV3101 C58C1 Rifr pMP90 Gmr, Koncz & Schell) in YEP in 250 mL baffle flasks. 2. Grow at
Yeast－Genomic－DNA－Prep
Grow 10ml YPD cultures o/n. Figure out cell density; inoculate 30 ml YPD and grow o/n so that cell density is approximately 2 x 108cells/ml the next
Streptomyces:Protocols/Spore－Prep
Spore Prep - Inoculating & Harvesting Description? A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored
porcine－brain－tubulin－prep
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
CSF－Extract－Prep－for－Spindle－Assembly
This protocol is essentially as described by Murray (1991), Cell Cycle Extracts. In Methods in Cell Biology, B.K. Kay and B. Peng, eds. (San Diego: A
Acid－Phenol－Yeast－RNA－Prep
This is the preferred method for yeast RNA preparationuse Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fres
Streptomyces:Protocols/MiniMaxi－Prep
Small Scale Plasmid Isolation (Mini / Maxi Prep)Description?A mini prep / maxi prep is used to isolate plasmid or cosmid DNA from bacteria, normally E
Establishment－of－Stable－Transfectant－of－CHO－Lec－Cells
Purpose and BackgroundsCHO lec 3.2.8.1 cellsCHO Lec 3.2.8.1 cells have four independent mutations in the N- and O- glycosylation pathways (Stanley, 19
CHO細胞表達體系及其特點
子生物學、分子免疫學等學科的發展使基因工程疫苗具有越來越重要的地位。在基因工程疫苗研究的動物細胞表達系統中，最具代表性的就是中國倉鼠卵巢細胞(Chinese Hamster Ovary，CHO)。它是用來表達外源蛋白最多也最成功的一類細胞。本文就 CHO細胞表達系統在疫苗研制中的應用做一綜述。C
TritonPrep－Method－for－bacterial－DNA－Purification
Triton-Prep Method for bacterial DNA PurificationGrow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube).Resus
A－quick－RNA－miniprep－for－Neurospora－mycelial－cultures
Most RNA isolation techniques currently in use have been developed for the processing of large quantities of material. These typically involve multip
PREP自動酶切儀使用方法
一 開機前檢查：1 檢查儀器臺面（DECK）上所有的實驗材料（Labware）。2 檢查SystemWater 水桶的水位。3 檢查恒溫循環水浴（Chiller）水箱的水位，并定期更換或填充Chiller 中的循環水。4 倒掉廢液桶中的廢液。二 開機步驟：打開Chiller、Heater、儀器及計算
CHO細胞無血清培養入門技術手冊
CHO細胞無血清培養入門技術手冊——深圳百恩維生物1、CHO細胞背景CHO細胞是中國倉鼠卵巢細胞(Chinese?Hamster?Ovary，CHO)，1957年美國科羅拉多大學Dr.?Theodore?T.?Puck從一成年雌性倉鼠卵巢分離獲得，為上皮貼壁型細胞，是目前生物工程上廣泛使用的細胞系。
CHO細胞的穩定轉染與基因表達
1、pcDNA3.1＋-gD的線性化: pcDNA3.1＋使用說明書推薦了以下幾個酶作為線性化酶(BglⅡ MfeⅡ Bst1107Ⅰ Eam1105Ⅰ PvuⅠ ScaⅠ SspⅠ),通過DNAMAN分析gD序列發現其帶有MfeⅡ酶切位點。2、轉染前一天,在60mm的dish中接種8×105個細胞
Quick－Yeast－DNA－Prep:－Isolation－of－Total－DNA－(genomic－and－plasmid)
Grow a 5 ml YPD O/N culture inoculated with a single yeast colony at 30 deg. Transfer culture to a small 13 x 100 glass tube. Spin down cells 2
CHO細胞染色體制備[Harvard－Medical－School]
Mitshison Lab, Department of Systems Biology, Harvard Medical Schoolhttp://mitchison.med.harvard.edu/protocols/chr1.htmlBuffers:Swelling Buffer (PME):
重組CHO細胞分離及蛋白收獲技術介紹
簡介：現今，通過哺乳動物細胞培養，新一代生物制藥得到了前所未有的發展。本文主要介紹通過中空纖維微孔過濾，以分離哺乳動物分泌蛋白的過程。起始濃度為2×105 cell/ml，細胞外蛋白產物是一種與白介素-2相似的10kD淋巴因子，是一種潛在的腫瘤治療藥物。該應用需要去除細胞和顆粒，而不裂解細胞，同時達
關于乙肝基因工程(CHO)疫苗的介紹
本疫苗系用基因工程技術將乙型肝炎表面抗原基因片段重組到中國倉鼠卵巢細胞(CHO)內，通過對細胞培養增殖，增殖分泌乙肝表面抗原(HBsAg)于培養液中，經純化加佐劑氫氧化鋁后制成，疫苗外觀有輕微乳白色沉淀。 1.接種對象 (1)乙肝易感者(表面抗原陰性，轉氨酶正常)。 ⑵用于阻斷母嬰傳播。給
島津Nexera－UC－Prep系統進入ANTOP專家評審階段
分析測試百科網訊 炎炎夏日，火的不僅有氣溫，火的還有這些分析儀器。經過廣大網友踴躍參與和熱情投票，島津申報的“制備超臨界流體色譜創新獎”已進入專家評審階段！島津Nexera UC Prep半制備超臨界流體色譜系統 島津Nexera UC Prep半制備超臨界流體色譜系統集合了Nexera UC
Fast－and－reliable－miniprep－RNA－extraction－from－Neurospora－crassa
We have developed a method for isolating high quality total RNA from?N. crassa?mycelia that reliably yields large quantities. It is possible to extrac
RNA－Extraction－(mini－prep):Trizol法實驗原理和步驟
RNA的制備與分析對于了解基因在轉錄水平上的表達與調控和cDNA的合成都是必須的，RNA的純度和完整性對于Northern blot，RT-PCR 和cDNA文庫的構建等分子生物學實驗都至關重要。RNA分離的方法很多，其中最關鍵的因素是盡量減少RNA酶的污染 實驗原理： ?Trizol 試劑
乙肝基因工程(CHO)疫苗的注意事項
(1)安瓿破裂、疫苗變質或有搖不散的塊狀物，不得使用。 (2)疫苗注射前要充分搖勻。 (3)接種疫苗時認明10ug/支及20μg/支兩種規格。 乙肝基因工程疫苗應于2-8℃條件下貯運，嚴防凍結，疫苗有效期為2年。
乙肝基因工程(CHO)疫苗的免疫效果介紹
乙肝基因工程疫苗自1992年獲得生產文號投入大量生產以來，免疫接種后安全可靠。血清學效果優于血源乙肝疫苗。兩種疫苗可以互相使用。對以前曾經用過血源疫苗未完成全程免疫的兒童，再用乙肝基因工程疫苗補充全程或加強免疫，同樣可以獲得滿意效果。
乙肝基因工程(CHO)疫苗的接種對象介紹
本疫苗系用基因工程技術將乙型肝炎表面抗原基因片段重組到中國倉鼠卵巢細胞(CHO)內，通過對細胞培養增殖，增殖分泌乙肝表面抗原(HBsAg)于培養液中，經純化加佐劑氫氧化鋁后制成，疫苗外觀有輕微乳白色沉淀。 接種對象 (1)乙肝易感者(表面抗原陰性，轉氨酶正常)。 (2)用于阻斷母嬰傳播。給