entire Protocols

Category: PCR Protocols: cDNA Synthesis Protocols

New RACE, a variation of RNA ligase-mediated-RACE (RLM-RACE) (Liu and Gorovsky 1993) departs from classic RACE (see 5'-End cDNA Amplification Using Classic RACE) in that an "anchor" primer is attached to the 5'-end of the mRNA before the reverse transcription step; hence the anchor sequence becomes incorporated into the first-strand cDNA if, and only if, the reverse transcription proceeds through the entire length of the mRNA of interest. - [Read 5'-End cDNA Amplification Using New RACE Protocol]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

This protocol is designed to adapt High Five™ (BTI-Tn-4) cells to serum-free suspension culture in HyQ SFX-Insect in 5 passages. The entire adaptation process takes approximately two weeks and the resulting adapted cells have doubling times of 16 and 20 hours and minimal clumping in suspension culture. - [Read Adaptation of High Five™ Cells to HyQ© SFX Insect Medium]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Shotgun sequencing of a large segment of DNA involves random fragmentation of the target region into smaller segments that are subsequently cloned into a bacteriophage M13 vector. The goal is to create a library of overlapping clones that provide at least fivefold coverage over the entire length of the target fragment. - [Read Generation of a Library of Randomly Overlapping DNA Inserts Protocol]

Category: Molecular Imaging

ImageJ - Image Processing and Analysis in Java. Useful program for MacOs MacosX and PC. Analysis includes: Measure area, mean, standard deviation, min and max of selection or entire image. Measure lengths and angles. Use real world measurement units s - [Read ImageJ - Image Processing and Analysis in Java]

Category: Microscopy Protocols: Electron Microscopy Protocols

The scanning transmission electron microscope precision and reproducibility of mass measurements are comparable with those of the analytical ultracentrifuge, the possibility of determining the mass not only of entire supramolecular assemblies but also of their distinct components has opened exciting new avenues which have occasionally been entered but are not yet fully explored. Includes: Principle and application (The GroEL:GroES complex). - [Read Imaging and Measuring Biomolecules & Their Assemblies by Scanning Transmission Electron Microscopy]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using bar1 Mutants Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using Low Cell Concentration Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: alpha-Factor Arrest Using Low pH Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Dilution of Stationary-Phase Cultures Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Hydroxyurea Arrest Protocol]

Category: Yeast Protocols: Yeast Cell Synchrony Protocols

It is sometimes desirable to have an entire population of yeast cells that is growing synchronously or is arrested at a unique point in the cell cycle. - [Read Inducing Yeast Cell Synchrony: Nocodazole Arrest Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for the isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. - [Read Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol]

Category: Microscopy Protocols: Confocal Microscopy Protocols

Confocal laser scanning microscopy (CLSM) is a relatively new light microscopical imaging technique which has found wide applications in the biological sciences. The primary value of the CLSM to the biologist is its ability to produce optical sections through a 3-D specimen-e.g., an entire cell or a piece of tissue - that, to a good approximation, contain information from only one focal plane. Article includes principle and applications of confocal laser scanning microscope. - [Read Looking Inside Cells and Tissues by Optical Sectioning with a Confocal Laser Scanning Microscope]

Category: PCR Protocols: PCR Cloning Protocols

Protocol exploits the discovery that Rnase A can efficiently cleave at single rC or rU bases embedded in double-stranded DNA.  Entire plasmid vectors are amplified using long, high-fidelity PCR with riboprimers, which carry a single rC residue at their 3' end.  Target DNA is amplified using similar primers, which also end in a rC residue. - [Read Ribocloning: DNA Cloning and Gene Construction Using PCR Primers Terminated with a Ribonucleotide]