"Transcriptomics & Functional Genomics"
"Promega GloMax- Multi+ Detection System"
The GloMax combined Photometer Luminometer and Fluorometer is located in room 2.058 corridor 3a. All users wishing to use of the instrument including BSc project students must have training before accessing this instrument and are encouraged to contact Dr K Laing. Training will be provided on a personalised adhoc basis along with on-going support as appropriate to the users needs.
What combined fluorimeters, plate spectrophotometers and luminometers are available for general use
- There are broadly at least three similar instruments covering this type of capability within Jenner Wing. These include the Biotek Synergy LX, BMG FluoStar Omega Lite and the Promega GloMax.
- The newest is the Biotek Synergy LX, combines luminesence, absorbtion (scanning between 200nm to 999nm similar to a naondrop but in a plate based system, it has a 0-4.0 OD dynamic range) and fluorescence (it has 510nm filter installed, so has limited scope for fluorescence unless additional filters have been added). It can take 6-384 well plates for absorbance 96 & 384 for all other modes see the Biotek web site for specifications. Note the instrument doesn't have the incubation module and runs off a halogen light source. Contact Daniel Meijles.
- Another nice instrument is the BMG FluoStar Omega Lite (contact Kim Jonas) located in the main lab in level 2 Jenner, its also quite new it has filters for Ex 355, 485, 544 and Em 460, 520, 590. In addition to those listed, we also have the filter sets for BRET and the TR module for TR-FRET. Standard luminescence can also be read, but this version note cannot read absorbance. It can also take 6-384 well plates reagent injection shaking and and incubation (see specifications for the Fluorstar OMEGA (noter this is not exactly the same as the Lite).
- The third instrument in this class is the older Promega GloMax which is more fully described below. This is located in the genomics lab level 2 corridor 3a. It has combined luminescence, and absorbance (6 fixed wave lengths see below) and Fluorescence: wavelength matched LEDs, with a top read PiN-photodiode wavelengths include UV (Ex:365nm, Em: 410-460nm & Ex:365, Em:515-570) , Blue (Ex:490, Em: 510-570nm), Green (Ex: 525nm, Em: 580-640nm), Red (Ex: 625nm, Em: 660-720nm) and AFC (Ex: 405nm, Em: 495-505 nm). More information generally about the GloMax see Promega's web page
Is there a charge and How do I access the GloMax
- There is no charge for access to this instrument. Electronic access to the room 2.058 on corridor 3a requires card access activation, in which case you need to fill out the access form you can down load it from here. Authorisation can be provided by Dr Ken Laing.
- Prior training on the use of the instrument is a requirement for access to the instruments and electronic calendars. This can be obtained by contacting Dr Ken Laing email@example.com
- Advanced Booking should be made, noting the time and duration you wish to use the instrument, giving your name and contact details. This can be done on-line using a Microsoft exchange calendar using Outlook or the OWA for the instrument.
- From time to time more information will become available on-line click here
How do I Book on-line using a Microsoft Exchange calendar
- With the shared Exchange calendar subscription is via invitation. To be added to the calendar, you should first contact Dr Ken Laing at firstname.lastname@example.org and arrange training as required and you will then be emailed an exchange calendar invitation.
- To do this you must use Outlook 2010. Outlook is available as a component of Microsoft Office.
- To install this calendar in the OWA, open the OWA and log in. You will see an email invitation to the calendar in your inbox.
- In the Outlook web app (OWA) the email will contain a "+" sign with adjacent text "ADD CALENDAR" this can be found just below the recipient list in the header, click this to add the calendar.
|Outlook Web App (OWA)
- To add in the calendar using the Microsoft Outlook 2010 email client open this application and log in. Where you will see an email invitation to the calendar in your inbox.
- Using Microsoft Outlook the email will have an exchange calendar icon at the top of the email next to the text "open calendar" click this icon to add the calendar.
- In each case the calendar will appear in the application on the left hand side when you select to display your calendars. A check box should allow you to display or hide the respective calendars as you wish.
- Select the check box "GloMax" calendar to see the current bookings on the machine. In the OWA these will be displayed overlayed by default, in Outlook you have to select this option.
- To add a booking in Microsoft Outlook, click on the time you would like to use the instrument, the tabbed calendar uppermost in the panel is the active calendar into which your booking will be inserted (for example in the image below this is for the CFX96 but the GloMax calendar would look similar). Enter your name and extn, press the carriage return to enter the booking.
- Now you may adjust the duration of the booking to the length of your run. To do this put your mouse over the bottom of the coloured block showing the booking you've just made and the cursor will change to a double ended arrow, click and drag to extend or reduce the duration. Your booking is complete. If you need drag and drop the booking to a different time if need be or select and delete the booking.
- Please do not book the machine for periods unreasonably longer than necessary, your run protocol will give you an estimate of this duration. If you have to cancel a run delete your booking and release the time for others to use. Please also dispose of your used plates and other rubbish!
How do I access my data?
- The GloMax can be operated either via a PC or the touch panel screen, it is easier to operate through the later. The data should be saved to the on board drive for temporary storage and can be retrieved using a USB flash drive or the PC.
What Wavelengths can I read
- Absorbance Visable: The instrument has an LED light source hand has 4 standard with two additional set filter holders that read in the range 405-750nm (405 [position A], 450, 560, 600, 690 [position B] & 750nm). It is has a photometric range of 5.0 OD units with a linear range of 0-4 OD units.
- Luminescence: Wavelength range 350-650nm with a detection limit 3x10-21 moles luciferase with a linear dynamic range of 8 logs.
- Fluorescence: wavelength matched LEDs, with a top read PiN-photodiode wavelengths include UV (Ex:365nm, Em: 410-460nm & Ex:365, Em:515-570) , Blue (Ex:490, Em: 510-570nm), Green (Ex: 525nm, Em: 580-640nm), Red (Ex: 625nm, Em: 660-720nm) and AFC (Ex: 405nm, Em: 495-505 nm) an additional kit is fitted. it has a detection limit of 1ppt of fluorescein and 6 logs linear dynamic range.
- Additional Specifications: Twin injectors 25-200ul in 5 ul increments. Linear and orbital shaking with variable speed up to 500 rpm. Heating from 2oC above ambient to up to 45oC.
- Kinetic feature using multiple plate reads is available in both Luminescent and fluorescent modes
What plate formats does the instrument require
The instrument is compatible with commonly used plate types and formats including 6, 12, 24, 48, 96 and 384 well plates. It can read wells at a single or multiple positions and reads in row order ie A1-12 followed by B1-12. For different plate formats a plate adapter must be used and orientated appropriately. 96 well plates should not be read with a lid The instrument uses a plate cover and this should be removed for all other plate types.
What Software is available
- The instrument comes with the Instinct software package for analysis, but data can be exported as either text or excel formats and most commonly people use excel or similar packages to analyse data.
More information generally about the GloMax see Promega's web page
- More detailed support and advice for this instrument can be obtained by contacting me Ken Laing email@example.com
- For more general information see my own web pages at http://www.ipc.nxgenomics.org/newsletter.htm and http://www.ipc.nxgenomics.org/
||Centre for Pathogen Genomics