It is important to remember, however, that if the assay tube remains in the. As you proceed through the assay, the. For instructions on the preparation of these standards, see the instruction manual that accompanies the assay you are using.
Standard 3 will appear on the screen. After Standard 3 is inserted, press GO. Note: It is important to prepare samples in the appropriate 0. After the calibration is complete or the values from the previous calibration have been accepted, insert a sample and press. The number displayed is the concentration of the nucleic acid or protein in the assay tube.
Calculate the concentration of your original sample see Calculating the Concentration of Your Sample, below Alternatively, choose Calculate sample concentration to have the.
Remove the sample from the instrument, insert the next sample, and press GO. If you did not choose Calculate sample concentration, calculate the concentration of your original sample by using the. These values can assist you in making a judgment regarding the performance of the. To minimize temperature fluctuations, store all kit reagents at room temperature and insert all.
Do not hold. This plot demonstrates that the curvefitting algorithm gives accurate values for quantitation. As such, we highly. Keep the tube. Often the problem is a relatively simple one that you can solve yourself with our direction. If it is determined by the Invitrogen Technical Service representative that the instrument should be returned for repair, a Decontamination Certificate will be faxed to you.
Once you have completed the certificate, signed it, and returned it to the Invitrogen. If you are in the United States, and the instrument is under warranty, Invitrogen will repair or replace the unit, and pay for return. Repair and shipping costs both ways are your responsibility if the instrument is. If you are outside the United States and have purchased the instrument from an authorized Invitrogen distributor, contact that. If it is. All rights reserved. These products may be covered by one or more Limited Use Label.
Licenses see Invitrogen catalog or www. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. For research use only. Not intended for any animal or human. Toggle navigation. Home Contact Us Sign In. Instruction Manual 20 Pages. Close If your hospital with access, please login via your intranet. Log in here About Us. Page 3 Table of Contents Inspection and Setup.
Position the instrument on a flat, dry surface. To the other end attach the correct plug adaptor, based on the electrical outlet configuration in your country.
Plug the power cord into the electrical outlet. Be sure to use only the power cord provided with your instrument. Powering the instrument with an unapproved power supply may damage the instrument.
The instrument is automatically powered on when it is plugged in. To recover from the sleep mode, press any of the four buttons on the instrument. Recovery from sleep mode returns the user to the same screen present when sleep mode commenced. After 10 hours of inactivity, the unit will power down automatically. Recovery from powered down mode returns the user to the Welcome screen.
You will be asked to supply the serial number, your name, and your contact details. Visit the web address below and follow the instructions. Do not hold the assay tubes in your hand before reading, as this will warm the solution and result in a low reading.
For greatest accuracy of the protein assay, the incubation time of the samples should be within 10 minutes of the incubation time of the standards. For this reason, if you want to perform multiple readings of a single tube, remove the tube from the instrument and let it equilibrate to room temperature for 30 seconds before taking another reading.
As you first use the instrument, you should perform a new calibration each time. As you become familiar with the assays, the instrument, your pipetting accuracy, and significant temperature fluctuations within your laboratory, you should determine the level of comfort you have using the calibration data stored from the last time the instrument was calibrated. Remember also that the fluorescence signal in the tubes containing standards and the samples is stable for no longer than 3 hours.
See Figure 2 in General Considerations for Analysis for an example of the calibration curve used to generate the quantitation results. Quantitation results are displayed directly on the screen for you to record. If data transfer is desired, you need to have: 1. As you proceed through the assay, the open spreadsheet will be updated with the data for standards and samples data transfer is initiated when the first standard is read.
If the fluorometer is powered down or in sleep mode, press any of the four buttons to power up the fluorometer. Note: If the instrument was in sleep mode, pressing any of the four buttons returns you to the last screen viewed before sleep mode commenced.
On the HOME screen, use the s or t key to highlight the assay of choice. Highlight either Run new calibration or Use last calibration and press GO. Note: When using the fluorometer for the first time, there will not be an option to Use last calibration. Note: It is important to prepare standards in the appropriate 0. Insert Standard 1 and press GO. Ensure you use the Standard 1 appropriate for the assay you are performing.
The reading will take approximately 5 seconds. Measuring Instruments Invitrogen Qubit 2. Fluorometer 41 pages. The Qubit fluorometer is a lab instrument developed and distributed by Invitrogen that, among other applications, is used for the quantification of DNA, RNA, and protein. The Qubit fluorometer uses fluorescent dyes to determine the concentration of either nucleic acids or proteins in a sample.
The other common method of measuring the concentration of nucleic acids and protein is the UV-absorbance method, which uses a spectrophotometer to measure the natural absorbance of light at nm for DNA and RNA or nm for proteins.
Because so many molecules absorb light at nm, this measurement is subject to inaccuracy due to potential contamination of the sample with these other molecules and is unable to distinguish between DNA, RNA, protein or free nucleotides or amino acids in the sample. The Qubit assays previously known as Quant-iT were developed and manufactured by the previous Molecular Probes now a part of Life Technologies. They have extremely low fluorescence until bound to their target molecule.
The difference in fluorescence between bound and unbound dye is several orders of magnitude. Upon binding to DNA, probably by intercalation between the bases, it assumes a more rigid shape and becomes intensely fluorescent. At a specific amount of the dye, the amount of fluorescence signal from this mixture is directly proportional to the concentration of DNA in the solution, even in the presence of other bio-molecules.
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