Judelson Lab Protocols

 

Dr. Judelson, University of California UC Riverside

 

 

 

Judelson lab protocols

 

DNA PREPS

• Alkaline lysis E. coli DNA preps

• E. coli boiling prep miniscreen

• Purifying DNA by PEG precipitation

• P. infestans genomic DNA

• Cleaning up genomic DNA using CTAB

• Quantitating DNA with Biorad fluorometer

RNA PREPS

• RNA extraction from P. infestans

CLONING AND MISC. DNA MANIPULATIONS

• Primer and oligonucleotide design strategies

• Restriction digestion of DNA

• Cloning and sequencing PCR products

• Advice on setting up ligations

• SSCP analysis

• Chemically competent E. coli cells

• Electroporation of E. coli

• Making electrocompetent E. coli DH5-alpha (Eppendorf protocol)

• Recycling electroporation cuvettes

• Plating lambda libraries

• Lambda plaque lifts

POLYMERASE CHAIN REACTION

• Primer and oligonucleotide design strategies

• Standard PCR (and colony PCR)

• RAPD analysis

• PCR amplification from phage plaques

MEDIA

• Amsco autoclave instructions

• General autoclaving guidelines

• Chinese media (Xu's)

• Lima bean media (ALBA)

• Formaldehyde (RNA) gels

• Rye and green pea media

HYBRIDIZATIONS AND GELS

• General information on DNA gels and photo documentation

• RNA gels (formaldehyde method)

• Northern blots using glyoxal

• cDNA probes from total RNA

• Preparing glyoxal for RNA gels

• Southern blotting of DNA (including making probes by random priming, and hybridization)

• Hybridizations using an oligonucleotide as probe

PHYTOPHTHORA TRANSFORMATION

• Phytophthora bombardment protocol

• Phytophthora electroporation protocol (zoospores)

• Protoplast transformation of P. infestans

MANIPULATIONS OF PHYTOPHTHORA

• Determining mating types

• Making zoospores and cysts

• Making crosses and germinating oospores

• Infecting leaflets with P. infestans

• Freezing P. infestans in liquid nitrogen

REPORTER ASSAYS

• GUS staining (histochemical)

• GUS assays (fluorometric)

MISCELLANEOUS METHODS

• Changing liquid nitrogen tanks

• Recovering bacteria from microtiter plate stocks

• G-force and rpm conversions for rotors in lab

• Removing photographic fixer stains from clothing

• How to use the Rainin Pipetman

• Rainin's Pipetman guide (includes replacement part numbers)

• How to keep a good laboratory notebook

• Advice on writing scientific papers

SAFETY

• Safety training document for lab

• Wipe test instructions

MISCELLANEOUS FORMS

• Radioactivity request and log sheet

• Log sheet for working in radioactivity area

• Log sheet for working in transfer hood