PlatinginTopAgar
1. Warm plates to room temperature before use. Cold plates causes the top agar to solidify irregularly. DO not warm plates to 37° as the top agar will take forever to solidify.2. Prepare top agar as the appropriate liquid medium with 0.7 agar. Keeping 100 mL bottles is convenient. For phages, use λ top agar, which is less rich and yields bigger plaques.3. Melt top agar in the microwave completely. Allow the agar to b......阅读全文
Plating－in－Top－Agar
1. Warm plates to room temperature before use. Cold plates causes the top agar to solidify irregularly. DO not warm plates to 37° as the top agar will
SOFT－AGAR－ASSAY－FOR－COLONY－FORMATION
Note: All volumes are calculated to cater for four plates per point.Base Agar1. Melt 1% Agar (DNA grade) in microwave, cool to 40 in a waterbath. War
Phage－Titer
IntroductionLambda phage is a commonly used vector for transgenes. Its very high rate of infectivity makes it ideal for creating large numbers of clon
Easy－Way－to－Clone－Genes－From－a－Phage－Library
Easy Way to Clone The protocol is oriented towards a C. albicans genomic library I made in Lambda Zap II on 7/97.The overall sequence of events is: •
Cell－counting/plating
OverviewCount the number of viable cells in a culture via dilution plating. The following assumes E. coli cells but it should apply to any type of cel
Pouring－Plates
1. For rich media, weigh out appropriate ingredients and place into a flask. Add water until appropriate volume. Use a flask at least 2 times larger t
Thawing－and－Plating－Cryopreserved－Hepatocytes
实验概要This protocol covers thawing and prep of cryopreserved hepatocytes for applications such as metabolic stability, intrinsic clearance, enzyme in
Bacterial－cell－culture
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
Noble－Agar－Assay
DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble
Preparation－of－Agar－plates
Prepare media and add 1.5 agar before autoclaving it (15g per liter). After autoclavation, cool the media in a 55 degree waterbath. Do not allow the s
Soft－Agar－Assay
Soft Agar AssayMake 0.6% media-agar mix for the bottom layer. To make 0.6% agar mix the following components (this makes 200 ml):2X DME 100 mlI
A－protocol－for－cleaning－and－reusing－the－large－25－x－25－cm－plates
We regularly reuse our large 25x25 cm plating trays; initially, however, we were plagued by gross microbiological contamination when reusing the trays
Agar－Plates－for－Selection－of－Clones－in－Bacteria
Cloning of PCR productsStocks:LB Agar: Luria Broth after Lennox:per LiterTryptone10 gYeast Extract5 gSodium Chloride5 gBact. Agar15 g pH 7.0Autoclave
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
Preparation－of－Phage－Lysates
Preparation of Phage LysatesInoculate 5 ml of lambda-broth in a glass culture tube with a single colony of an appropriate host strain of E. coli. Incu
Human－Embryonic－Stem－(ES)－Cell－Protocols——Matrigel－Aliquoting－and－Plating
Aliquoting Matrigel:Day one:Put the sterilized tip box (either 200 ml or 1000 ml tips), sterilized eppendorf tube container, and appropriate pipettor
Preparing－Antibiotics－Stock－Solution－and－Ampicillin－Agar－Plates
AMPICILLIN Beta-lactam-antibiotics are not very stable when dissolved. Slow but steady degradation happens even when frozen to -20°C. Therefore comme
Preparing－Lambda－DNA
Preparing Lambda DNA1- Coliphage lambda DNA is a widely used vector for recombinant DNA. The middle third of its 48,000 bp contains no genes required
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
Dropout－plates－for－yeast
Materials(Solutions are all available from the media room)200ml bottle of 2x SD200ml bottle of 4% agar -- make sure to sign it out40% glucoseCSM minus
沉淀反应实验：琼脂扩散(agar－diffusion)实验
琼脂扩散实验琼脂扩散是抗原抗体在凝胶中所呈现的一种沉淀反应。抗体在含有电解质的琼脂凝胶中相遇时，便出现可见的白色沉淀线。这种沉淀线是一组抗原抗体的特异性复合物。如果凝胶中有多种不同抗原抗体存在时，便依各自扩散速度的差异，在适当部位形成独立的沉淀线，因此广泛地用于抗原成分的分析。琼脂扩散实验可根据抗原
peg6000－agar为什么会有沉淀
沉淀是肯定的因为PEG本来就与琼脂不相溶，它与黄原胶、海藻酸钠和明胶也不相溶只能与化学结构相似的PVP或者PAM等少数高聚物互溶（PVA都不行），请酌情参考。另外PEG-6000因为分子量高羟值低它溶于热水没问题但是一冷却下来就会析出的，从PEG-2000开始就是分水岭了……
Probiotic－Attributes－of－Lactic－Acid－Bacteria－Isolates
IntroductionPresently the consumption of probiotic food is highly popular worldwide for their health beneficial effects. Commonly used probiotic bacte
重组DNA的分离、克隆与测序实验手册6
Electroporation ProtocolPreparation of Electro-competent Cells:1. Grow XL1-Blue cells on a tetracycline plate (20 ug tet/ml of LB agar)2. Inoculate 3
Spheroplast－Transformation
MaterialsYPD platesYPD1 M sorbitol 182 g/l (Sigma S7547)2 M sorbitol 36.4 g/100 mlSCE (per liter)1 M sorbitol (182 g)100 mM citric acid trisodium salt
cDNA－LIBRARY－SCREENING
PREPARE SOLUTIONS1. 10mM MgSO4, 0.2% Maltose LB (100 mL):Mix 1.0 g of Bacto-Tryptone, 1.0 g of NaCl, 0.5 g of Yeast Extract, and 1.0 mL of 1M MgSO4. A
Screening－Bacterial－Colonies－Using－Xgal－and－IPTG:－αComplementation
实验概要 α-complementation occurs when two inactive fragments of E. coli β-galactosidase associate to form a functional enzyme. Many plasmid
In－vitroculture－of－early－chick－embryos
1. Use sterile technique. Prewarm Howard's Ringers in petri dish andagar/albumin culture dish to 37ºC.2. Crack 2-day egg into large sterile petri
Glycerol－Agar－（甘油琼脂）的成分和适用范围
Glycerol Agar （甘油琼脂）Peptone （蛋白胨） 5g Beef extract （酵母膏） 3gGlycerol （甘油） 20g Top water （自来水） 1000mlAgar （琼脂） 15g pH 7.0-7.2
Cosmid－Cloning:－Cell－preparation,－DNA－packaging,－and－Cell－Transfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man