Construction of S. pombe MicroArray containing promoter regions
We developed the
S. pombe Promoter MicroArray in collaboration with the Södertörns University College in Sweden and the Max Planck Institute in Berlin, Germany.
Up to 4976 specific primer pairs have been designed and synthesized to amplify promoter regions of
S. pombe ORFs.
We added a universal TAG sequence of 14 and 15 bases to all forward and reverse primers respectively. The two-step method of PCR reaction generates C6-amino modified PCR products with an average length of 500 bp.
Spotting design
The
S. pombe promoter MicroArrays are manufactured by printing PCR amplicons suspended in optimized spotting buffer for high coupling efficiency of DNA to the most consistent aldehyde coated glass slides. This spotting chemistry provides us the highest hybridization intensities, which is a consequence of a better binding of the probe to the slides and their accessibility to the cDNA targets.
We use ChipWriter™ Pro (Virtek) which is a high precision dispensing robot to spot the PCR products onto aldehyde glass slides. The robot is designed to collect 100 nl of DNA solution and to deposit 0.6 nl per spot. For
C. albicans MicroArray, we use simultaneously 32 pins to place the spots in area of 3.6 x 1.8 cm. In order to generate high quality spots, the printing procedure is performed under a tight controlled humidity and temperature environment. This allows having the complete control on the spot morphology, the spot diameter and uniformity. This configuration of the robot generates 32 blocks, each containing 20 x 20 dots with 200 microns spacing center to center with a spot diameter of 100 microns.
| Name |
Quantity |
Reference |
EUR |
|
| S. pombe promoter MicroArrays |
5 |
AR-IGRP-01 |
1900.00 |
|
Quality Control
The
S. pombe promoter MicroArrays are certified to generate a high quality data.
All printed slides are previously selected for coating uniformity and low intrinsic background.
The MicroArrays are visually inspected for spotting quality (doughnuts), uniformity, integrity and for perfect alignment of spots and grids.
From each batch of printed MicroArray, three are randomly chosen for further quality control. We use the Terminal Transferase reaction to assess the amount of DNA covalently attached to the slides. Finally, we perform a real hybridization using labelled cDNA obtained from heat-choc cultured cells.
Delivery times
10 WD
Packaging
The MicroArrays are shipped in a light-tight box and they can be stored at room temperature for the minimum of 3 months without loss of signal intensities.
You will receive an Excel™ file containing the list of
S. pombe promoters and their coordinates in the MicroArray.
The package includes comprehensive protocols for RNA extraction, cDNA synthesis, and labelling and hybridization conditions that give reproducible and high-quality hybridization results.
Shipping conditions
Room temperature
Storage conditions
Room temperature