微生物学技术

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 ...

Protocol for quick tests of restriction enzyme activity in novel species Belavin P. A. Dedkov V.S. Degtyarev S.Kh. A method to detect restriction endonucleases in bacterial colonies. (1988) Applied Bi ...

Getting The Most Out Of Your BugsNative lysis is a staple protocol in practically every biochemistry lab yet there is significant variability in the procols I have read. The intention is to liberate t ...

OverviewThis is a quick and easy yeast colony PCR protocol that does not require zymolyase step. Updated Protocol: Blackburn Lab: Quick and Easy Yeast Colony PCR v2.0 MaterialsStandard PCR machine tub ...

Intergeneric Conjugation and OverlayDescription Transfer of plasmid/cosmid DNA from a host strain e.g. E.coli ET12567 to the recipient strain e.g. Streptomyces coelicolor. Approx. Duration: Conjugati ...

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.coli. An overnight culture of cells is lysed and t ...

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 (ssD ...

Description Transform E.coli cells with plasmid/cosmid DNA using the method of electrophoration (inserting plasmids into E.coli). Approx. Duration: Preparation ~30 minutes Making the competent cells ~ ...

SummaryCleaner stocks of T7 that concentrates and purifies T7 bacteriophage. ProtocolGrow 100mL of permissive cells to a density of 108 to 109 cells/ml at 30°C in a rotary shaking water bath. Inoculat ...

Materials50mL 7H9 mycobacterial medium + 3mL per transformation 102mL 10% glycerol (possibly a few mL more if you are doing many transformations) 400mL PBS + 0.1% Tween 80 per transformation electropo ...

Spore Prep - Inoculating & HarvestingDescription A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 20% sterile glycerol at -20°C. It will last for man ...

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. ProtocolAdd 5mL fresh overnight (BL21 if wild-type ...

大多数豆科植物与真菌共生。丛枝菌根真菌（AM）对磷（P）的吸收和根瘤菌对氮（N2）的固定具有重要的农学和生态学意义。植物-AM真菌-根瘤菌三个共生如何高效吸收营养的机制受到很多关注。AM真菌和根瘤菌能够有效地增加固氮和植物对土壤中磷的吸收，但这破坏了根部阴阳离子平衡，过多的阳离子需要从根部分泌出来才能达到平衡，H+是分泌的最主要阳离子。然而，无论根瘤能够直接释放H+引起土壤的酸化， ...