The Electronic Protocol Book---A Quick and Practical Guide for Biologists (original) (raw)

Antibody (37)

Absorb antibody with acetone protein powder

Antibody idiotype determination

Antibody storage

Antibody-Protein A or G binding

Coupling of antibody to biotin

Coupling of antibody to protein A or G beads

Coupling of GST-protein to glutathione beads

Coupling of peptides for antibody production

Coupling of peptides to Sepharose beads

Dissolving peptides

ELISA --- alkaline phosphatase detection

ELISA --- horseradish peroxidase detection

ELISA plates

Estimation of antibody concentration

Fluorescein labeling of proteins (antibodies) (I)

Fluorescein labeling of proteins (antibodies) (II)

Immunoprecipitation

Peptide antibody design

Prepare paraformaldehyde fix buffer

Production schedule --- monoclonal antibody

Production schedule --- polyclonal in chicken

Production schedule --- polyclonal in goat and sheep

Production schedule --- polyclonal in rabbit

Production schedule --- polyclonal in rodent

Purification --- affinity (I)

Purification --- affinity HiTrap (II)

Purification --- ammonium sulphate precipitation

Purification --- chicken IgY

Purification --- phospho-antibody

Purification --- protein A column

Purification --- protein G column

Staining --- ABC (DAB)

Staining --- cells on slide

Staining --- formalin-fixed section

Staining --- frozen section

Staining --- membrane

Staining for immunohistochemistry or FACS

Apoptosis (3)

Extraction of low MW DNA fragments

TUNEL

TUNEL --- FACS

Appendix (41)

Amino acid structures

Centricon spin speed and time

Centrifugation froce conversion

Classification of amino acids

Codes for degenerate nucleotides

Common lab stock solutions and molar concentration

Common PCR and sequencing primers

Commonly used agarose gel DNA markers

Comparison of embryonic development stages

Consensus sequences for protein kinases

Dye migration in agarose and polyacrylamide gels

Extinction coefficient for oligonucleotide and protein

Gene nomenclature guidelines

Genetic code --- human mitochondria

Genetic code --- universal

Genome sizes and number of genes

Hydrophilicity index of amino acids

Kozak translation initiation sequence

Loading buffers

Mail DNA/Clone/Stock samples

Male and female symbols

Microcon cutoff

Molecular weight for amino acids

Molecular weight for oligonucleotide and protein

Nucleoside and nucleotide structures

Oligo melting and PCR annealing temperature

Peptide and phosphodiester bond

Periodic table of the elements

pH indicators

Phosphate buffer chart

pK values of amino acids

Polyadenylation signal

Promoter sequences

Protease inhibitors

Proteases

Protein localization signals

Single-letter amino acid codes

Splicing signal

T3/T7/SP6 minimum promotor sequences

Units

Weight-mole conversion for DNA and protein

Bioinformatics (51)

3D structure database of macromolecules

Automation --- automatic archiving old files

Automation --- Microsoft Excel macros

Automation --- Microsoft Excel tables

Automation on Mac --- AppleScript

Automation on Mac --- iDo Script Scheduler

Automation on PC --- batch files

Automation on PC --- scheduled task

Blast

Blast --- local

Capture screen image

Converting to pdf files

Drawing chemical structures

Dos and Unix commands

EST assembler

Evolutionary trees

FASTA format

GenBank identifier syntax

Gene annotation databases

Hidden Markov Model --- search for remote relatives

HTML

HTML forms

Human disease gene database

Install and run Java

Java JAR applications

Mass spectrometry

Metabolic pathway database

Multiple sequence alignments

ORF finder and gene finder

Perl

Perl Example 1 --- file preview

Perl Example 2 --- remove digits and spaces

Perl Example 3 --- insert links

Perl Example 4 --- count frequency

Perl Example 5 --- data merge

Perl Example 6 --- archive old files

Perl On Mac

Perl On Mac --- MacPerl

Perl On Windows

Pfam --- database of protein domains

Protein secondary structure prediction

PubMed

Shading of multiple sequence alignments

SNP database

Translation

UniGene --- non-redundant gene clusters

VecScreen --- vector sequence contamination

Viewing postscript files

Visulization of 3D structures

Visulization of 3D structures on Mac

Web restriction digestion

C. elegance (1)

Worm protocols

Cell Biology (45)

Alkaline phosphatase activity --- secreted

Adaptation to new media

beta-galactosidase acitivity assay --- 96-well

Cell count using Coulter Counter

Cell count using hemocytometer

Cells on slides --- adherent cells

Cells on slides --- suspension cells

Chloramphenicol Acetyl Transferase (CAT) Assay

Chromosome spread

Cloning by FACS

Cloning by limiting dilution

Epitope tags

Establish stable cell clones

Establish stable cell clones --- 96-well

Establish stable cell clones --- cloning cyclinders

Establish stable cell clones --- drug concentrations

Freezing cells

Heat inactivation of serum

Luciferase activity assay --- 96-well

Metabolic labeling of proteins

MOI/pfu/TCID50

Mycoplasma cure

Mycoplasma test --- PCR

Mycoplasma test --- visulization of DNA

ONPG beta-galactosidase activity assay

Phalloidin staining of filamentous actin

Preparation of poly-L-lysine-coated slides

Quick freezing of cells on plates or flasks

Serum starvation

Soft agar assay of colony formation

Sterilization techniques

Sub-culture of adherent cells

Sub-culture of suspension cells

Thawing frozen cells

Tissue culture waste

Transfection --- Calcium phosphate

Transfection --- electroporation

Transfection --- liposomal

Transfection --- non-liposomal

Transfection --- suspension cells

Trypsin/EDTA

Visualization of DNA --- DAPI

Visualization of DNA --- dye list

Visualization of DNA --- Hoechst

X-gal staining of cells

Cell Cycle (9)

BrdU labeling of S phase cells

FCAS analysis

Histone kinase activity assay

Synchronization of cells --- double thymidine block

Synchronization of cells --- growth condition

Synchronization of cells --- mitotic shake-off

Synchronization of cells --- nocodazole

Synchronization of cells --- summary

Synchronization of cells --- yeast alpha-factor

Cloning (74)

3'-RACE

5'-RACE

Agarose gel electrophoresis

Annealing of oligonucleotides

Antibiotic stocks

Bacterial glycerol stock

Bacterial host strains

Bacterial media

Bacterial plates

Bacterial plates --- X-gal

Calcium chloride competent cell

Common sources of cloning contaminations

Deletion of DNA/RNA --- ethidium bromide staining

Dephosphorylation of DNA --- CIP

Dephosphorylation of DNA --- SAP

Detection of DNA --- fluorometer (Hoechst )

Detection of DNA --- fluorometer (Pico green)

Detection of DNA/RNA --- methylene blue staining

Detection of DNA/RNA --- spectrometer

Detection of DNA/RNA --- SYBR Green staining

Detetion of DNA --- colorimetric

Detetion of DNA in PAGE --- UV shadowing

DNA concentration measurement

DNA ligation

DNA ligation (quick ligation)

Effective range of separation of DNAs in agarose gels

Effective range of separation of DNAs in PAGE

Effective range of separation of DNAs in PAGE/Urea

Ethonal precipitation of nucleotic acids

Growing bacterial cells --- 96-well

Patch grids for 100 mm plates

Pcik a single clone

Plasmid miniprep --- alkaline lysis

Plasmid miniprep --- boiling

Plasmid prep --- 96-well

Plasmid prep --- CsCl

Plasmid prep --- Midi/Maxi column (Qiagen)

Plasmid prep --- Qiagen mini

Polyacriamide gels

Polyacriamide/urea gels

Purification of DNA from agarose gels --- DEAE cellulose

Purification of DNA from agarose gels --- Qiagen column

Purification of DNA from agarose gels --- silica beads

Purification of DNA from PAGE gels

Replicate clones in 96 well plates (I)

Replicate clones in 96 well plates (II)

Restriction digestion

Salmon sperm DNA

Screening for insertion --- blue-white (lacZ)

Screening for insertion --- plasmid size (I)

Screening for insertion --- plasmid size (II)

Sequencing --- ABI Big Dye Terminator

Sequencing --- Dideoxy Chain Termination

Sequencing of GC-rich DNA

Sequencing troubleshooting

Southern --- Chemiluminescence detection

Southern --- radioactive probe

Southern --- stripping

Southern transfer

Stab culture

Stab stocks

Sterilization techniques

Streaking

Terminal deoxynucleotidyl transferase

TE-saturated Phenol

Transfer of nucleotic acids to membrane --- capillary

Transfer of nucleotic acids to membrane --- large fragment

Transfer of nucleotic acids to membrane --- PAGE

Transfer of nucleotic acids to membrane --- vacuum

Transformation

Transformation --- competent cells

Transformation --- electroporation

Transformation --- electroporation competent cells

Vector for TA cloning

Fly (17)

Anesthetizing flies

Collecting eggs

Culture stocks

Distinguishing males from females

Establish clean cultures from dechorionated eggs

Fly resources

Fly traps

Food recipe from Bloomington stock center

Genetic nomenclature for Drosophila melanogaster

Instant Fly Food

Mating

Quarantine

Single-fly DNA prep for PCR

The fruit fly life cycle

Virgins

X-gal staining

Yeast paste

High throughput drug screen (9)

HIV reverse transcriptase activity assay

How to calculate IC50

IC/ED/GI/LD

Measure cell proliferation --- 3H-labeled thymidine

Measure cell proliferation --- BrdU ELISA

Measure cell proliferation --- sulforhodamine B

Measure cell proliferation --- tetrazolium

Measure cell proliferation --- XTT

NCI/NIH drug screen program

Histology (15)

Antigen retrieval --- autoclave

Antigen retrieval --- microwave

Blood smear

Counterstain

Fixation solutions

Formaldehyde/paraformaldehyde/formalin

Giemsa stain

Gram stain

Mounting medium

Mounting medium --- anti-fade

Paraffin embedding

Sectioning --- frozen tissue

Sectioning --- paraffin block

Tissue fixation --- ethanol

Tissue fixation --- formalin

Lab equipments (16)

Autoclave spore test

Centrifuge

Electroporation --- parameters for different cells

Electroporation --- recycle cuvettes

Homogenizers

Hybridization oven

Laminar flow hoods and chemical fume hood

pH meter

pH paper

Shakers

Spectrometer

Storage freezer

Vacuum

Water

Water bath

Liquid scintillation counter

Labeling of probes (21)

DNA end-labeling

DNA labeling by nick translation (32P)

DNA labeling by nick translation (biotin/digoxigenin)

DNA random labeling with 32P-dCTP

DNA random labeling with Biotin/Digoxigenin

Ethanol precipitation of DNA probes

Incoporation efficiency --- filter binding assay

Incoporation efficiency --- TCA precipiation

Intensifying screen

Isotopes used in biological sciences

Labeling of cDNA --- Amino-allyl dye coupling

Labeling of cDNA --- Cy3/Cy5 direct

Labeling of DNA --- Amino-allyl dye coupling

Labeling of DNA in agarose gel

Labeling PCR products with Biotin/Digoxigenin

Liquid Scintillation Counter efficiency

Liquid Scintillation Counting

Oligonucleotide mole/weight/concentration calculations

Purification

Purification --- PAGE

Purification --- Sephadex column

Labeling of proteins (8)

32P labeling of phosphorylated proteins

35S labeling of protein in bacteria

35S labeling of proteins

Coupling to biotin

Coupling to fluorochromes

In vitro translation

Iodination

Iodination --- IODO-BEADS

Library (10)

Amplification of helper phage

Bac/PAC/cosmid/fosmid prep --- 96 well

PCR amplification of library insert

Phage clone --- pick clones for storage

Phage clone --- storage

Phage DNA prep --- lambda phage

Phage library --- tittering

Phage library screening --- PCR (I)

Phage library screening --- Southern

Prepare host cells for infection

Microarray (13)

Data analysis --- filtration

Data analysis --- merge

Data analysis --- missing data fill-in

Data analysis --- normalization

Hybridization

Labeling of samples

Microarray --- Hierarchical clustering

Microarray --- Tree view

Microarray Explorer

Microarray platforms

Sample labeling

Software and data mining

Prepare slides for printing

Miscellaneous (13)

Bacteria smear

Buffer exchange

Buffer exchange --- Desalting column

Buffer exchange --- dialysis tubing

Buffer exchange --- Slide-A-Lyzer Dialysis Cassettes

Concentrating samples

Concentrating samples --- centricon

Lyophilization --- freeze-dry

Mouth Buccal swab

Mutagen Ames test

Mutagen Ames test --- automation

Proteasome overlay

Wet mount

Mouse (12)

Animal husbandry

How to tell male from female

Isolation of genomic DNA from mouse tails

Mouse diet --- NIH open formula

Mouse ear punch numbering system

Mouse resources

Mouse/Rat diet suppliers

Routes of injection in mouse

Sex of mouse embryo and cell line

Staining of mouse embryo --- AP

Staining of mouse embryo --- lacZ

The mouse life cycle

PCR (17)

Asymetric PCR

PCR

PCR --- DpnI-mediated mutagenesis

PCR --- long products

PCR amplification of cDNA library inserts

PCR amplification of single clones

PCR amplification of yeast clones

PCR cleanup --- 96-well

PCR cleanup --- agarose gel

PCR cleanup --- Microcon

PCR cleanup --- Qiaquick spin column

PCR cleanup --- SAP/ExoI

PCR construction of insertion/deletion/mutation/fusion

PCR from blood smears

PCR primer design

PCR random mutagenesis

Sequencing of PCR products

Protein (32)

Acetone protein powder

Degenerate sequences --- decoding from protein

Drying of polyacrylamide gels --- membrane

Drying of polyamcriamide gels --- vacuum

Effective range of separation of proteins in PAGE

In vitro transcription/translation --- TNT

In vitro translation --- Rabbit Reticulocyte Lysate

PAGE --- gel casting

PAGE/Native

PAGE/Native --- recipe calculator (30% stock)

PAGE/Native --- recipe calculator (40% stock)

PAGE/SDS --- recipe calculator (30% stock)

PAGE/SDS --- recipe calculator (40% stock)

Protein concentration --- Bradford (I)

Protein concentration --- Bradford (II)

Protein concentration --- estimate by staining

Protein concentration --- Lowry

Protein concentration --- Modified Lowry

Protein concentration --- UV

Protein precipitation --- acetone

Protein precipitation --- ethanol

Protein precipitation --- TCA

Protein purification for micro-sequencing --- PVDV

Staining of proteins

Staining of proteins --- Coomassie blue

Staining of proteins --- quick Coomassie blue

Staining of proteins --- silver

Staining of proteins ---SYPRO

Western --- dot blot

Western --- ECL

Western --- stripping

Western transfer

Protein expression (20)

Baculovirus --- virus stock storage

Baculovirus --- amplifying virus stock

Baculovirus --- cluturing of insect cells

Baculovirus --- freezing and thawing of insect cells

Baculovirus --- infection

Baculovirus --- plaque assay

Baculovirus --- plaque purification of virus stock

Cleavage --- Factor Xa

Cleavage --- thrombin

Concentrating virus (PEG)

Expression of protein in bacterial cells

Freezing virus

Protein refolding

Purification of GST-fusion protein

Purification of his-tagged protein --- HiTrap

Purification of his-tagged protein --- TALON

Purification of inclusion body proteins

Purification of inclusion body proteins --- HiTrap

Purification of moltose-binding protein fusion

Screen of recombinant protein expression

Recipes (1)

Recipes

Report genes (6)

Alkaline phosphatase (AP)

beta-galactosidase (lacZ)

Chloramphenicol Acetyl Transferase (CAT)

Enhanced Green Fluorescent Protein --- Living Colors

Horseradish peroxidase (HRP)

Luciferase activity assay

RNA (19)

DEPC treated RNAse-free solutions

Formaldehyde-Agarose gels

In vitro transcription

Northern

Polyadnylation --- Ambion PolyA tailing

Reverse transcription

Ribosomal RNA sizes

RNA purification --- acid phenol:chloroform extraction

RNA purification --- DNase I digestion

RNA purification --- guanidinium thiocyanate

RNA purification --- hot phenol

RNA purification --- LiCl precipitation

RNA purification --- polyA (I)

RNA purification --- polyA (II)

RNA purification --- Qiagen Rneasy

RNA purification --- Trizol

RNAse protection

RT-PCR --- one-step

siRNA design

Safety and Waste (10)

Bacterial and cell culture waste

Blood

Cleaning of mercury spills

Common harzardous materials in biology lab

Decontamination of ethidium bromide solutions

MSDS (Material Safety Data Sheet)

Radiation protection

Radioactive waste

UV protection

Wipe test for beta emitting radioisotopes

Signal transduction (3)

Peptide substrates for kinases

Protein dephosphorylation

Quantitate kinase activity --- cellulose phosphate filter

Statistics (7)

Basic statistics

Chi-square test

Linear regression

Normal distribution

Outliers

Statistics software

Student t-test

Yeast (15)

Beta-gal filter assay

Disrupting yeast cells

Genetic nomenclature for yeasts

Liquid sporulation

Phalloidin staining of filamentous actin

RNA prep --- phenol (I)

RNA prep --- phenol (II)

Spheroplast

Transformation --- electroporation

Transformation --- LiAc (I)

Transformation --- LiAc (II)

Transformation --- spheroplast

Yeast glycerol stocks

Yeast media

Yeast resources