DNA基础知识

1. Constitutive - those enzymes that are continuously synthesized in the cell (e.g. the enzymes of glycolysis or the lac repressor protein); 2 In ...

b-Galactosidase Activity Assay -- Marian Price-Carter 9/7/00Day 1: Start overnight cultures in assay medium.Negative control: cells lacking b-galactosidase such as LT2; positive control: cells with hi ...

CAT Assay Ref. Seed and Sheen Gene 67 271-277 (1988)Normal reaction is Chloramphenicol + Acetyl-CoA - Acetylchloramphenicol Assay uses butyryl-CoA (more hydrophobic) 14C-Cm+ Bu-CoA-Bu-14C-Cm Km (Cm) ...

InroductionSingle-cell electroporation (SCE) is a technique we have developed to deliver genes into individual cells within intact tissues although it may also be applicable to cells in disperse cultu ...

CAT ASSAY 1. Transfer cells to a 15 ml tube. 2. Add 5 ml TBS- to flasks shake & pour into tubes. 3. Spin down the cells @ 1k rpm for 5'. 4. Resuspend in 1 ml TBS- . 5. Transfer to 1.5 ml tubes. ...

CAT Enzyme Assay System With Reporter Lysis BufferChloramphenicol acetyltransferase (CAT) encoded by a bacterial drug-resistance gene inactivates chloramphenicol by acetylating the drug at one or both ...

CAT ASSAY (liquid phase) 1. Transfer cells to a 15 ml tube. 2. Add 5 ml TBS- to flasks shake & pour into tubes. 3. Spin down the cells @ 1K RPM for 5'. 4. Resuspend in 1 ml TBS-. 5. Transfer to 1. ...

β-Galactosidase Enzyme Assay System with Reporter Lysis Bufferβ-Galactosidase is a commonly used reporter molecule. The β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer (Cat.# E2000) is ...

β-Galactosidase Enzyme Assay System with Reporter Lysis Bufferβ-Galactosidase is a commonly used reporter molecule. The β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer (Cat.# E2000) is ...

Establishment of Stable Transfectant of CHO Lec Cellsby Jun Takagi 6/15/2000Purpose and BackgroundsCHO lec 3.2.8.1 cellsCHO Lec 3.2.8.1 cells have four independent mutations in the N- and O- glycosyl ...

1. Remove gel slice contain DNA fragment and place in 10 volumes of:300 mM NaOAc pH 7.0 300 ml 1 M NaOAC pH 7.01 mM EDTA 2 ml 500 mM EDTA pH 8.0698 ml ddH202. Incubate at 22 C for 30 min. Transfer gel ...

OUTLINE OF PROCEEDINGS USED IN TISSUE CULTURE TRANSFECTIONS1. Remove the required cells from liquid nitrogen and thaw at 37°C.2. Add the thawed cells to 10mls C2GM that has been equilibrated in the 3 ...

OUTLINE OF PROCEEDINGS USED IN TISSUE CULTURE TRANSFECTIONS1. Remove the required cells from liquid nitrogen and thaw at 37°C.2. Add the thawed cells to 10mls C2GM that has been equilibrated in the 3 ...

Electroporation of P. aeruginosa Preparation of Electrocompetent Cells Inoculate a 5-ml of L-broth with cells of the P. aeruginosa strain to be electrotransformed. Grow culture overnight at 37°C on a ...

Electroporation of P. aeruginosa Preparation of Electrocompetent Cells Inoculate a 5-ml of L-broth with cells of the P. aeruginosa strain to be electrotransformed. Grow culture overnight at 37°C on a ...

Materials:SOB medium E. coli host strain such as DH5αWBtRNA5 M ammonium acetate100% ethanol70% ethanol0.5X TESOC mediumtransformation platesI. Preparation of E. coli cells for electroporation.1. Use a ...

How to Make Competent Cells Step 1: Take an 100 mL aliquot of frozen cells from the -80oC and innoculate about 500 ml to 1 L sterile LB broth. DO NOT ADD antibiotic since these cells do not have an pl ...

Deproteination using phenol/chloroform (Maniatis et al. 1982)'Phenol' as used is Analar grade. Phenol should be melted at 65°C8-hydroxyquinoline added to a final concentration of 0.1% and equilibrated ...

Dialysis (using dialysis tubing)Molecularpourous membrane tubing is used for desalting protein and DNA isolation / purification. It comes in several diameters and pore sizes (for molecular weight conv ...

NH4Ac and EtOH precipitation of DNAAdd NH4Ac (10 M stock or solid) to the sample for a final concentration of 2.5 M mix (spin at 4 C transfer the supernatant to a new tube; optional spin for extra pur ...