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1. Initial inoculum:- From solid media, touch an isolated colonies with a sterile applicator or toothpick.- From liquid media, touch the culture with the end of a sterile applicator.- From a frozen culture, scrape the surface with a sterile applicator; you need not take visible clumps.- From a stab tube, insert a sterile applicator into the agar and withdraw; do not take agar chunks.2. Draw the inoculated end of the ......閱讀全文
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
The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" CellsJoseph SambrookPeter Maccallum Cancer Insti
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
Growth and storage of Agrobacterium tumefaciensStrain GV3101: resistant to gentamycin and rifampicin so add 25-50 ug/ml Gentamycin, 10 ug/ ml rifampic
Simple and Efficient Method (SEM) to Make Competent Cells Preparation of Frozen Competent of DH5α 1) 250 ml TB solution 10mM Pipes or 1
2.2 Interaction mating - large scaleWith a few modifications, the procedure described above can be used to test for interactions between a single prey
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 1
Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei
ConclusionsQuorum-sensing signaling systems involving the interaction between a signaling peptide and its cognate histidine kinase receptor are widely
Metabolic biotinylation of mammalian cell receptors for imagingBakhos A. Tannous , btannous@hms.harvard.edu, Massachusetts General Hospital and Harvar
1B. Cloning 1. A caveat on dephosphorylation: the most common reason for failure to obtain colonies is a
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
實驗概要 Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at l
In the polymerase chain reaction (PCR), a thermostable DNA polymerase amplifies DNA that is flanked by known sequences. The known sequences correspond
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and reci
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and reci
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt
DNA抽提（主要內容如下）· Working with DNA· DNA Extraction from Bacteria and Other Organisms· DNA Extraction f
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polym
Desalt DNA template by EtOH precipitation in NaOAc followed by at least 2x washes with 70% EtOH. Resuspend in 5 - 15 μL of sterile H2O.Rinse cuv
6. Simultaneous digestion of the pUC vector with both enzymes in the presence of 3 units of Shrimp Alkaline Phosphatase (
實驗概要 α-complementation occurs when two inactive fragments of E. coli β-galactosidase associate to form a functional&nbs
Need 1.5-2 x 107 cells from a 2 day culture.1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold
Flagella and motilitymonotrichous flagella - the bacterial cell has a single flagellaperitrichous flagella - the bacterial cell has several
Flagella and motilitymonotrichous flagella - the bacterial cell has a single flagellaperitrichous flagella - the bacterial cell has several
ProcedurePrepare serial 10-fold dilutions of transformed bacteria in LB and spread 100 μL onto LB/Amp plates, as described in the Electroporation prot
Genomic DNA libraries Size of some genomes and chromosomes:Comparative Sequence Sizes(Bases)(yeast chromosome 3)350 ThousandEscherichia coli (bac
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
Three transformation systems have been reported for the rice blast fungus Magnaporthe grisea (Parsons et al. 1987 Proc. Natl. Acad. Sci. USA
Differential Interference Contrast (Nomarski, DIC, Hoffman Modulation Contrast)PrincipleDifferential interference microscopy requires several optical