Streptomyces:Protocols／PCR
Description Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template double stranded DNA (dsDNA) to single stranded DNA (ssDNA). Custom designed primers/oligonucleotides then anneal to their target sequence on the ssDNA. The DNA polymerase then extends the primers in a 5’ → 3’ direction, creating a complementary double strand. The cycle ......閱讀全文
Streptomyces:Protocols/PCR
Description?Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template doubl
Streptomyces:Protocols/Conjugation
Intergeneric Conjugation and OverlayDescription?Transfer of plasmid/cosmid DNA from a host strain, e.g. E.coli ET12567 [pUZ8002], to the recipient str
Streptomyces:Protocols/Transformation－by－Electroporation
Description?Transform?E.coli?cells with plasmid/cosmid DNA using the method of electrophoration (inserting plasmids into?E.coli).Approx. Duration:Prep
Streptomyces:Protocols/Spore－Prep
Spore Prep - Inoculating & HarvestingDescription?A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 2
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
LCM－PROTOCOLS
Slide SectioningParaffin blocks-?For?DNA?analysis:Special LCM processing schedule is followed.The water bath is cleaned using RNAse Zap?, rinsed thoro
CGH－Protocols－（三）
Hybridizationreagents:?labeled tumor and normal-DNA (see protocol Nick translation)?salmon sperm DNA, 10 mg/ml (e.g. Promega)?human Cot1 DNA, 1 mg/ml
CGH－Protocols－（一）
Metaphase chromosome preparationMaterials:?RPMI 1640 medium?fetal calf serum (FCS), 20%?Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best
Smolke:Protocols/Western
OverviewBlotting for large V5-tagged proteins in?S. cerevisiaeMaterialsY-PER (Pierce)Halt EDTA-free Protease Inhibitor (Pierce)NuPAGE Novex Bis-Tris 4
Neutralizing－Bioassay－Protocols
Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri
CGH－Protocols－（二）
DNA preparation by cryotom tissue dissectionPreparations/Materials:?Cool cryostat down to -20 to -30°C about 3 hours prior to dissection?Label eppendo
DAPI－Counterstaining－Protocols
實驗概要The ?blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; ?it appears to associate with AT clusters in the minor groove. Binding
General－Cloning－Protocols
Large Scale Preps:?(See Large scale plsasmid prep protocol for more details)Cultures: Inoculate a 5 mL LB/Amp (50 - 100 μg/mL) culture in early a.m. w
CGH－Protocols－（四）
CGH Image acquisitionImages were acquired through a Zeiss Axiophot fluorescence microscope using a Plan NEOFLUAR oil objective x63, N.A. 1.25 (Zeiss,
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
Rat－Blood－Collection－Protocols
實驗概要The procedure presented below describes a method for collecting rat blood.實驗步驟Rat should be fully anesthetized (e.g., unresponsive to toe pinch).1
ORNL－MICROARRAY－HYBRIDIZATION－PROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5:A. MATERIALSRNeasy? Mini Kit (Qiagen; Cat # 74106)?SuperScript II RT (200U/μL) (Life Technologies; Cat #
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis?I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA.?EmbeddingB.?CuttingC.?StainingII. Pr
FISH－protocols－for－Drosophila2
?3.?Methods3.1 RNA Probe Preparation1.?? Different strategies can be used to prepare template DNA for synthesizing antisense RNA probes by?in vitro?tr
DataONE:Protocols/Find－GEO－reuses
Identify reuses of GEO datasetsAimThe aim of this protocol is to collect data on the reuses of datasets in the published literature. This particular p
Red－Blood－Cell－Lysis－Protocols
實驗概要BioLegend’s ?Red Blood Cell (RBC) Lysis Buffer (Cat. No. 420301) has been designed, ?formulated, and tested to ensure optimal lysis of RBCs in sin
FISH－protocols－for－Drosophila1
.1 RNA Probe Preparation?(see?Note 1)1.?? 1.5 mL microcentrifuge tubes or standard 96-well V-bottom microplates.2.?? RNAse free water.3.?? T7, T3 or S
Standard－Protocols－Autoradiography－(35S)
Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in
Slice－and－Explant－Culture－Protocols－–－Hevner－lab－2002
for axon tracing & cell culture studies in vitro using embyros age E11.5 – E16.5modified from Rubenstein lab and Price lab protocols1. Setupa. 2 hr be
Human－Embryonic－Stem－(ES)－Cell－Protocols——Embryonic－Bodies
Let human ES cells grow until the colonies are large and the cells are pretty piled up - about the time when you would normally split or even a day pa
Human－Embryonic－Stem－(ES)－Cell－Protocols——Media－and－Reagents
Serum Free Media for human ES cells on MEFs:?can last for 7-10 daysFinal ConcentrationAmount for 250ml Stock solution80% DMEM-F12200ml20% KO Serum Rep
Protocols－for－the－preparation－of－tumour－cells－for－s.c.－injections－in－mice
For the establishment of solid tumour in nude mice, tumour cells are treated in a sterile environment under a cell culturing hood. Disposable sterile
Protocols－for－the－preparation－of－tumour－cells－for－s.c.－injections－in－mice
For the establishment of solid tumour in nude mice, tumour cells are treated in a sterile environment under a cell culturing hood. Disposable sterile
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
Human－Embryonic－Stem－(ES)－Cell－Protocols——Freezing－Human－ES－Cells
?Collagenase cells for approximately 7 minutes at 37 °C (until edges of colonies are curling up).With a 5 ml pipet, gently pipet and scrape colonies f