实验基础

OverviewGeneral guidelines on how to grow up a culture of Acetobacter xylinum ATCC strain 53582 . Preparation of Acetobacter MediaTo prepare ∼500 ml of liquid Acetobacter media add the following: Gluc ...

MaterialsGlass culture tubes with metal caps and labels Growth medium from media room or customized Glass pipette tubes Parafilm EquipmentVortexer Fireboy or Bunsen burner Motorized pipette Micropipet ...

IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids BAC) into a bacterium. Bacterial cells into which foreign DNA can be transformed are called competent. Some bacteria ...

AbstractChIP-Chip stands for Chromatin Immunoprecipitation and chip in the sense of DNA microarray. It is a technique to determine the genome-wide binding sites of a DNA-binding protein. While the bas ...

MaterialsPCR SuperMix High Fidelity VF2 primer (5''-TGCCACCTGACGTCTAAGAA-3'') VR primer (5''-ATTACCGCCTTTGAGTGAGC-3'') Deionized sterile H2O strip of PCR tubes 2-log DNA ladder 0.8% E-Gel® from Invitr ...

OverviewThis is a method to assemble two BioBricks using the Clontech In-Fusion PCR Cloning Kit and maintains BioBrick standard formats. There are currently several BioBrick assembly standardsthat all ...

Solutions/reagents:LB broth + selective marker50% sterile glycerolTEG (25mM Tris-Cl 10mM EDTA 50mM dextrose)20 mg/ml lysozyme10% SDS4M NaOHautoclaved waterSolution 3 (3M potassium-acetate 2M acetic ac ...

IngredientsIngredients are per culture; make enough for one extra culture to allow for pipetting error). 150μL sterile 50% glycerol 1mL TEG (25mM Tris-Cl 10mM EDTA 50mM dextrose) 111μL 20mg/mL lysozym ...

BackgroundThis protocol is adapted from "Molecular Cloning: A Laboratory Manual" Second Edition Sambrook Fritsch and Maniatis. It is a quick inexpensive way to purify large numbers of plasmids (I used ...

MaterialsFor purifying plasmid DNA from Escherichia coli cells the Qiagen Spin Miniprep Kit produces quite reliable results. Do not autoclave solutions containing isopropanol or MOPS; use sterile filt ...

plasmid miniprep All ''miniprep'' methods reported so far for the isolation of plasmid DNA involve multiple pipetting extraction centrifugation and changes of minifuge tubes. For screening large numbe ...

OverviewGeneral guidelines for the electro-transformation of Lactobacillus sake as described by Berthier et al. and as used by Alegre et al. with Lactobacillus plantarum. Materials50 ul of L.plantarum ...

OverviewElectrotransformation procedure for Lactobacillus acidophilus ProcedurePrepare Electrocompetent cells Inoculate overnight culture at 10^6 CFU/ml in MRS containing 1% glycene Harvest at early-l ...

OverviewGeneral overview and guidelines on how to grow up a culture of Lactobacillus MaterialsMRS broth (difco) MRS agar (difco) anaerobic conditions ProcedureFollow instructions to make MRS plates an ...

OverviewThis page details a electrotransformation protocol for Lactobacillus bacteria specifically Lactobacillus delbruckii subsp. bulgaricus and Lactobacillus delbruckii subsp. lactis. The protocol i ...

OverviewInstructions on how to prepare Lactobacillus plantarum competent cells before electrotransformation. MaterialsMRS media Culture of L. plantarum cells MgCL2 (10mM) sucrose glycerol Centrifuge c ...

Tn5 Transposase productionTn5 transposase is the key enzyme in forming transposomes for random transposon insertions. It is sold by Epicentre at ridiculously high price. Here we make it from a plasmid ...

Materialsovernight culture of Mesoplasma florum in ATCC 1161 medium preparing 10 ml cultures with 1 10 100 and 1000 μl of infected cultures assures that one of these will be ready the next day at the ...

Choice of EnzymeHutchison used DraI as a cutter but this is a blunt cutter making religation difficult MboI cuts at GATC sites and is insensitive to 5-me dCTP methylation (sensitive to methylation of ...

Transposome DNA construction from plasmid cuttingCut the transposome out of the containing plasmid with PvuII enzyme which cuts at the correct location at the mosaic end. Cut also with an enzyme which ...