Protein Expression Eukaryotic cells A. thaliana

Get tips on using pANC232 (xylF) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylF

Get tips on using pANC231 (xylE) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylE

Get tips on using pANC230 (xylD) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylD

Get tips on using pANC223 (xylC) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylC

Get tips on using pANC210 (xylB) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylB

Get tips on using pANC209 (xylA) to perform Protein Expression Prokaryotic cells - A. cellulolyticus xylA

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

Get tips on using pBYR2T-EGFP to perform Protein Expression Prokaryotic cells - A. tumefaciens GFP

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.