PreparingOvernightBacterialCulture
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 10 gstir to dissolvepH to 7.0autoclave to sterilizecool to 50-60 degrees Cadd antibiotic if neededAgar plate with bacterial colonies:scrape bacteria glycerol stock such as E coli DH5 (LifeTechnologies) or scratch a single colonies from an old agar plate with bacteria col......阅读全文
Preparing－Overnight－Bacterial－Culture
Materials:Sterile LB medium (Luria-Bertani Medium) with or without antibiotic:water 500 mlbacto-tryptone 10 gbacto-yeast extracts 5 gsodium chloride 1
Bacterial－cell－culture
MaterialsGlass culture tubes with metal caps and labelsGrowth medium, from media room or customizedGlass pipette tubesParafilmEquipmentVortexerFireboy
细菌培养
Preparing Overnight Bacteria Culture (LaboratoryExperiments.com)This is a basic procedure for high school students and useful for those who are new to
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
Cosmid－DNA－Isolation
实验概要Isolation of high yields of highly pure cosmid DNA using PureLink™ HiPure Plasmid Purification Kits.实验原理The PureLink™ HiPure Plasmid Purification
Preparation－of－Bacterial－Proteins－for－Analysis－by－2DPAGE
The following protocol has been developed for preparing soluble bacterial proteins in a form suitable for analysis by 2D-PAGE. The procedure was princ
Long－Term－Storage－of－Bacterial－Strains
Purpose:Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40 glycerol. It is lab policy to
E.Z.N.A.™－Fastfilter－Plasmid－Mega－Protocol
实验概要The E.Z.N.A.™ family of products is an innovative system that radically simplifies extraction and purification of nucleic acids from a variety
Midiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
质粒的小量制备
· Standard (alkaline lysis) Mini-Prep (Goldberg Lab)Standard protocol for mini-prep and recipe for solution I, II and III.Alkaline Lysis Minip
Growing－Overnight－Cultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-ca
E.Z.N.A.®－Plasmid－Maxi－Kit－vacuum－Protocol
实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c
Expression－Protocol－in－M9－Minimal－Media－via－T7－Promoter
The following protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 min
Preparing－cells－and...
实验概要The method provides a protocol and tips for BrdU staining in tissue sections.Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic
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
Preparing－a－Selenomethionyl－Protein
PurposeThe protocol describes how to prepare selenomethionyl (Se-Met) protein using a regular E.coli strain. Selenium can be used for phase determinat
Expression－Protocol－in－M9－Minimal－Media－via－T7－Promoter
Expression Protocol in M9 Minimal Media via T7 PromoterThe following protocol has been used successfully to 15N or 13C/15N label our proteins using ou
Transformation－of－E.－coli－by－Electroporation
实验概要 Electrocompetent bacteria are prepared by growing cultures to mid-log phase, washing the bacteria extensively at low temperature, and
E.Z.N.A.®－Plasmid－Maxi－Kit－Spin－Protocol
实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c
Preparation－of－Plasmid－DNA－by－Alkaline－Lysis－with－SDS:－Minipreparation
实验概要Plasmid DNA is isolated from small-scale (1-2 ml) bacterial cultures by treatment with alkali and SDS.主要试剂Buffers and Solutions: Alkaline lysis
Large－Scale－Plasmid－Preps:－Qiagen/Cesium－Method
Most plasmids can be adequately prepped by kits containing DNA binding columns. These columns do not do a great job of separating plasmid DNA from con
Differentiating－Neural－Stem－Cells－into－Neurons－and－Glial－Cells
实验概要The protocols in this section describe the steps involved in differentiating neural stem cells (NSC) to neurons, astrocytes, and oligodendrocyte
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
超级感受态细胞的制备
The Inoue Method for Preparation and Transformation of Competent E. coli: "Ultra Competent" CellsJoseph SambrookPeter Maccallum Cancer Institute and T
Streaking－Bacterial－Stocks
Principle:Bacterial cells are streaked onto a medium to obtain an independent isolate. This is done to reduce the likelihood of working with a culture
Preparing－chemically－competent－cells
MaterialsPlate of cells to be made competentTSS bufferLB mediaIceGlassware & EquipmentFalcon tubes500μl Eppendorf tubes, on ice200ml conical flask200μ
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
Lambda（噬菌体）DNA－Miniprep
David HarryInstitute of Forest GeneticsUSDA Forest ServicePacific Southwest Research StationAugust 26, 1993Background :There are many published method
DNA纯化手册2
Key points to observe: a. Use a endA1- E. coli strain for plasmid propagation and isolation whenever possible. The instability of plasmids isolated fr