Services - Flex-Chips

Custom Microarray Services from TeleChem International, Inc. provide unprecedented access to the microarray revolution at an unbeatable price. By uniting the power of microarray technology with the universality of the Internet, TeleChem makes microarrays affordable, versatile and easy to use. Every microarray is manufactured in one of our state-of-the-art class 100 cleanroom facilities, with the most advanced robotics, surfaces and coupling chemistries in the industry.  Our particle- and ribonuclease-free manufacturing laboratories provide microarrays of dazzling quality. Flex-Chips allow customers to retain confidentiality of their samples by allowing the customers to prepare and send samples of their choice in a 384 well format. arrayit.com then prints the customer samples on optically flat www.arrayit.com/Products/Substrates/SuperAmine, SuperAldehyde, SuperEpoxy, SuperProtein or other surface. The result is a virtually unlimited variety of microarray biochips for biomedical, pharmaceutical and agricultural research applications. Because the customer prepares the samples, the all results are completely confidential. Imagine any molecule, from any organism, for any application on a pristine surface and you are beginning to understand the breadth and depth of our Custom Microarray Services. TeleChem leads the way in custom microarray manufacturing.

Please send requests for quotes by electronic mail to arrayit@arrayit.com.

Table of Contents

• Introduction
• Quality Control and Use
• Description of Services
• Technical Assistance
• Short Protocols
• Complete Protocols
• Equipment Requirements
• Publications
• Tips for Design and Use
• Product Contents
• Ordering Information
• Storage Conditions
• Warranty

Introduction
Congratulations on taking a big step towards improving the affordability, quality and speed of your genomics, biomedical, pharmaceutical and agricultural research. This booklet contains all the information required to take full advantage of TeleChem's Flex Chips.

Quality Control and Use
TeleChem International, Inc. takes every measure to assure the quality of our Flex Chips.  The finest microarray cleanroom technology was used to develop these products. Rigorous quality control monitoring guarantees that each microarray biochip conforms to the highest industry standards.  Please be aware that certain types and uses of microarrays may be patent protected. TeleChem International, Inc. does not endorse the improper use of its Custom Microarray Services.  Please consult your intellectual property expert to assure that your particular application is licensed properly.

Description of Services
Flex Chips from TeleChem International, Inc. provide a virtually unlimited variety of microarray biochips at an affordable price.  All of our microarrays are fabricated in a state-of-the art class-100 cleanroom, with 0.1 µm filtered air and precise temperature and humidity control.  This nearly particle-free, sterile setting provides a custom microarray manufacturing environment that rivals facilities in the semiconductor industry.  Users will appreciate the following product features:

• Customer provides the samples (up to twenty-five 384 well microplates)
• Pristine, optically flat glass printing substrates
• Proprietary, covalent coupling chemistries that are stable to 100°C
• Substrates (25 x 76 x 0.96 mm) fit all commercial microarray detection systems
• Substrate corner chamfer enables unambiguous slide orientation
• Barcoding allows automated chip identification
• Any sequence, any organism, any application
• Arrays of DNA, RNA and peptides
• Utilize a wide range of parallel analysis applications
• Examine both hybridization and protein interactions
• State-of-the-art class-100 cleanroom sets a new standard for microarray fabrication
• Robotics with ± 1 µm positional accuracy
• Triplicate* spotting assures high data precision and low CVs
• TeleChem's advanced Stealth printing technology assures highest microarray quality
• Total confidentiality of all results!
• Develop new assays, publish, discover and patent results
• Internet savvy services leverage electronic design and ordering

*Replicates spotted depend on the number of samples printed. Larger number of samples may be double spotted or single spotted please inquire for more details.

Reasonable turnaround time of 1-2 weeks (larger orders may take longer).

Technical Assistance
Please contact us if you have any comments, suggestions, or if you need technical assistance. By electronic mail: arrayit@arrayit.com (under the subject heading please type "ArrayIt® technical assistance"). By telephone: (408) 744-1331, Monday—Friday PST 9:00am - 4:30pm. Please remember that we want to hear about your successes!

Short Protocol (Flex-Chips)
1. Determine the number of 384 well plates to be arrayed and number of substrates to be printed. The Flex Chip pricing table at the end of this section will help you in determining the approximate pricing. To order please email us at arrayit@arrayit.com. Once the quote is issued, a 50% down payment or a PO number (credit approval may be necessary, please see Ordering Information below) is necessary before the Flex Chips services can proceed.
2. arrayit.com will send you the appropriate number of 384 well plates with foil seals, environmental proof shipping bags and appropriate TeleChem spotting solution.
3. Re-suspend samples in TeleChem’s Micro Spotting Solution or Micro Spotting Solution Plus or Protein Printing Buffer (see Table 1) and transfer 10 µl per well to the 384 well microplates.
4. Seal 384 well microplates with foil covers (seal every well completely!).
5. Mail sealed microplates back to TeleChem.
6. Receive printed and bar-coded Flex-Chips (2-6 weeks depending on the details of your order).
7. Process the Flex Chips to remove unbound target sequences.
8. Label and purify probes.
9. Perform a hybridization or biochemical reaction.
10. Wash Flex Chips to remove unbound probe molecules.
11. Detect the fluorescent signals.Quantify and model the data.

Table 1. Sample Preparation Guide for Flex-Chips.

Complete Protocols (Flex-Chips)
1. Calculate the total cost of the order using the data provided in Ordering Information below, the per chip cost is dependent on the number of chips ordered and the number of elements printed. For example, if the customer has one 384-well plate of samples and wishes to obtain 25 printed chips, the per chip cost is $107 for a total cost of $2,675. The customer would then send a PO number or down payment for 50% of the order referencing the quote number to: TeleChem International, 524 East Weddell Drive, Sunnyvale, CA 94089. Ph: (408) 744-1331, Fax (408) 744-1711 or by email to arrayit@arrayit.com. Upon receipt of the order and PO number or down payment, arrayit.com will send the customer the required number of 384 well plates, foil plate covers, information labels and TeleChem Micro Spotting Solution (MSS) or Micro Spotting Solution Plus (MSS Plus).  The 50% down payment is required for all orders to defray the upfront costs associated with completely custom work. 

2. Upon receipt of the plates, re-suspend samples in 1X Micro Spotting Solution or Micro Spotting Solution Plus as described in Table 1. Make sure samples are in the appropriate concentration and are re-suspended completely. Samples that are mixed well give superior results in printing. All samples must be suspended in the 1X MSS or MSS Plus provided by TeleChem. arrayit.com is not responsible for poor printing quality that results from the use of sample buffers other than 1X MSS or MSS Plus, or poor quality results due to inappropriate concentrations of material.

3. Transfer 10.0 µl per well to provided 96 or 384 well microplates provided by TeleChem. Use only these plates, customer samples sent in the wrong plates will be returned for re-plating!

4. Seal the 96 or 384 well microplates with the foil covers provided such that every well is sealed completely. This is done manually by pressing down on the foil cover over each microplate well. If properly sealed, samples are stable against dehydration for >30 days at room temperature. This allows users to prepare a large number of samples over a period of days or weeks for simultaneous shipping. arrayit.com is not responsible for improperly sealed plates. Dehydrated or desiccated samples will be returned to the customer for re-plating.

5. Fill out the information labels provided then freeze plates at -20°C overnight prior to shipping.  All samples should be shipped frozen to prevent sample leakage

6. Mail sealed and frozen microplates to TeleChem International, Inc. at 524 East Weddell Drive, Sunnyvale, CA 94089. Place plates inside a box with dry or wet ice to maintain appropriate temperature and include packing materials (bubbles or peanuts) to minimize agitation.

7. Receive printed and bar-coded Flex-Chips after a period of 2-6 weeks, customer samples will be returned with the printed Flex Chips. Once your plates are received we will provide a more exact timeline. The time to process orders depends on the volume of orders in the queue and on the size of your order. TeleChem International, Inc. will make every attempt to process your order as quickly as possible.  Microarrays made from 1 384-well microplate are printed in triplicate with 4 pins at 4.5 mm center-to-center spacing for a total array size of 9 mm x 9 mm. Microarrays printed with 2-25 microplates are printed in triplicate with up to 48 pins at 4.5 mm center-to-center spacing for a total array size of up to 18 mm x 60 mm. Large numbers of samples may require a larger printing area and less than triple spotting. Flex-Chip shipping schedules vary depending on the volume or orders. The shipping time of your Flex Chips will be estimated upon receipt of you samples. 

8. Obtain Flex Chips shipped directly to you by Express Mail or by the shipping method that you specify.  All Flex Chips will arrive complete with barcodes and corner chamfer. The barcode contains an identifying number that is unique to each chip in your order.  The corner chamfer is provided to allow the user to determine easily which side of the substrate contains the printed microarray and defines the top of the substrate.  The printed microarray is located on the same side of the substrate as the barcode.  The corner chamfer should appear in the upper right corner if the barcode is at the bottom of the substrate. You will also receive a content file that shows the location of each printed target sequence on the microarray substrate.

9. Process the Flex Chips to remove unbound printed array elements.  Processing protocols vary according to the Substrate used and the target sequences. Detailed protocols are available electronically on the SuperAmine and SuperAldehyde product page. 

10. Label and purify probes. There are a myriad of enzymatic and synthetic methods used to label probes derived from RNA, oligonucleotides and proteins. Samples can be labeled using direct labeling whereby fluorescent tags are incorporated into the molecules in the sample. Alternatively, samples can be labeled by indirect means prior to hybridization such that epitopes such as biotin or amino allyl are incorporated into the sample, then the molecules are made fluorescent using fluorescently labeled streptavidin or reactive fluorescent dyes.  Post hybridization detection can also be achieved using secondary labeling molecules such as dendrimers (Genisphere, Inc) or antibody-containing fluorescent molecules. Samples can be nucleic acids, proteins or other types of molecules.  Users should select the labeling procedure that works best for their application.  Labeling procedures and tips are available under DNA Microarray Protocols, Super Microarray Substrates, Hybridization Products, and Fluorescent Probe Purification.  An electronic library containing nearly 1,000 publications provide a wealth of information on labeling and purification procedures. Our specialists are happy to provide advice and feedback on probe labeling and purification.

11. Perform a hybridization or biochemical reaction. Probes should be reacted with the processed Flex Chips to allow the molecules to interact with their cognate targets on the chip.  This step is best accomplished by reacting the sample to the processed microarray underneath a cover slip positioned over the microarray.  The cover slip should extend ~2 mm beyond the edges of the microarray on all sides to ensure that the sample covers all the spots. Sample volumes are typically ~2.0 µl per cm², such that total reaction volumes of 3.0 µl and 20 µl work well for the 12 mm x 12 mm and 22 mm x 40 mm cover slips, respectively. The corner chamfer should be at the upper right side of the substrate relative to the user when the microarray is held vertically (with the barcode at the bottom). Buffers suitable for nucleic acid hybridizations include 5X SSC + 0.2% SDS, 1X UniHyb^TM and 1X HybIt^®.  Other types of buffers such as those containing SSC, SSPE, SDS, and TritonX-100 have also been used successfully. Sample is best added to the microarray by placing the drop at the edge of the cover slip and allowing the sample to spread out between the microarray and the cover slip by capillary action.  Once the sample is added, immediately place the microarray in an enclosed chamber such as a Hybridization Cassette and bring the reaction up to temperature as quickly as possible.  It is critical to prevent desiccation during the biochemical reactions, as drying is the major source of elevated background fluorescence in microarray experiments.  Desiccation is prevented by adding 5-25 µl of buffer to the reaction chamber. Reaction temperatures are typically 37-65°C, though different experiments may require higher or lower reaction temperatures.  Reaction times are usually 2-12 hours.  Information on hybridization reactions can be obtained under Hybridization Products, Super Microarray Substrates, and DNA Microarray Protocols.  Our specialists are happy to provide advice and feedback on hybridization, protein, and other types of biochemical reactions involving Flex Chips.

12. Wash Flex Chips to remove unbound probe molecules. Once the Flex Chips are reacted with labeled probe samples, the chips must be washed to remove unbound molecules. Following the biochemical reaction, unbound sample should be washed off the surface. For cDNA hybridizations, the following protocol works well.  Remove the microarray from the Hybridization Cassettes and place it in a High-Throughput Wash Station and wash according to the protocols provided with the Microarray Wash Buffers.  Wash conditions should be modified for different types of reactions and different applications.  Make sure the cover slip falls off within 1 minute after transfer into the wash buffer. Wash buffers pre-heated to 42°C can be used to speed the release of the cover slip from the microarray.  Failure to remove the cover slip within 1 minute after transfer to the wash buffer may lead to elevated background fluorescence.  There are numerous washing protocols available and details are available under DNA Microarray Protocols, Super Microarray Substrates, Hybridization Products, and Microarray Wash Buffers.  An electronic library containing nearly 1,000 publications provide a wealth of information on microarray wash procedures. Spin the substrates dry by centrifugation for 10 seconds in a Microarray High-Speed Centrifuge.  Our specialists are happy to provide advice and feedback on Flex Chip washing strategies.

13. Detect the fluorescent signals.  Once the Flex Chips are reacted and/or stained, and washed, the fluorescent signals must be detected.  Detection is best performed using one of the high-quality commercial instruments available.  Commercial systems are available from PE (Packard Bioscience), Axon, Applied Precision and several other vendors. Detection should be performed such that the most intense signals on the microarray produce sub-saturating values. Rare species in the probe mixture can be quantified using instrument settings that maximize detectivity. Adjusting the laser power, PMT gain, capture time and other parameters provide enormous detectivity in microarray assays.

14. Quantify and model the data. Data quantification and mining can be performed using a variety of custom and commercial software packages.

Equipment Requirements
· Fluorescence microarray detection device (Packard Biosystems ScanArray, Axon, Virtek ChipReader etc.)
· Hybridization cassettes (TeleChem)
· High-Throughput Wash Station (TeleChem)
· Glass cover slips (many vendors)
· Quantitation software (BioDiscovery ImaGene, Packard Biosystems QuantArray)

Publications
A comprehensive database of nearly 4000 microarray publications is available electronically at http://arrayit.com/e-library/.

Tips for Design and Use
Table 2. Many different types of Flex-Chips are possible. The different types allow the user to explore a vast array of molecular interactions and a multitude of applications.