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| If you are first user or would like to use this service, please contact: |
| For more details about pricing and collaborations please contact: |
| - High Throughput Colony and Plaque Picking |
| - Gridding |
| - Replicating |
| - High-Density Arraying |
| - Re-arraying |
| - Micro-Array Analysis |
High-Throughput Colony and Plaque Picking |
| This technique is used for the construction of cDNA, genomic libraries or
multiple copies of sequences of interest. Pieces of DNA or cDNA are inserted into a vector
(e.g. plasmid, cosmid, phage, plaque) and then incorporated into a host system (e.g. yeast
or bacteria). A dilution of the library is then spread onto an agar plate and grown
overnight into discrete colonies. The colonies of interest are then selected by their
growth characteristics, (e.g. size, color, roundness) using the image processing system.
These colonies are picked from the agar plate into a microtitre-plate with growth media
containing the appropriate antibiotics (to select for clones containing the insert and to
prevent contamination) and glycerol (to aid survival during repeated freeze/thawing). The
microtitre plate containing the clones is then grown at the appropriate temperature
overnight. |
| Gridding (also Spotting) |
| This technique is used to identify the piece of DNA or cDNA of interest
within a library. The library is gridded from the microtitre plates onto nylon membranes.
The DNA is then fixed to the nylon by cross-linking with UV light. The membranes are
exposed to a radioactive or fluorescent labeled probe, and washed to remove any unbound
probe. The membrane is imaged and scored to identify the positive microtitre plate
wells with the DNA/cDNA of interest. To increase the number of copies of the DNA/cDNA, the
polymerase chain reaction (PCR) is used. The product is then extracted and cleaned up
before sequencing. |
| Replicating |
| This is the production of multiple copies of a library. It is recommended
the source microtitre plates (384 well) are used only once to produce replicates to
prevent cross-contamination and death of clones due to repeated freeze-thawing. |
| High-Density Arraying/Micro-Arraying |
| Same as gridding, but onto glass slides at an increased density of from 4
to 8 x384 well plates per slide. This produces slides that represent from 1456 - 2912
different samples. |
| Re-Arraying |
This technique has two main functions:
1. To re-arrange a fully identified library sequentially;
2. To select specific target wells and re-arrange them into another set of plates for
further analysis, e.g. to remove false positives (also known as subtraction) |
| GeneTAC® Array
Analysis |
| Automate the imaging and analysis of gene micro-arrays Quick image
acquisition of micro-arrays spotted on standard microscope slides Dual fluorescent label
detection Automatic imaging of up to 24 slides at a time Powerful analysis tools for rapid
identification, quantification and reporting of spots. Reporting features include plate
identification, location and matches to public domain, commercial or your database.
Compact, bench top design Complete system for spotting, hybridizing and analyzing
micro-arrays when combined with FlexysTM robotic workstation, Genomic IntegratorTM and the
AutoHybridizerTM Flexys Librarian software facilitates re-arraying and subtraction of your
DNA library |
| Info |
Unit |
Robot time |
| Colony and Plaque Picking |
24 colonies |
2 minutes |
| Gridding on paper/cellulose membranes |
384 spots |
2 minutes |
| Replicating |
1384 well copy |
2 minutes |
High-Density Arraying on slides
Re-arraying |
384 spots |
10 minutes |
| Micro-Array Analysis |
1 slide |
~20 minutes |
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Plant
Biotechnology and Genome
Center