1
Q
explain how to start amplification with the qiagility
A
- normalize samples and PCs in elution (pointed) tubes
- make master mix in an elution tube with an overage of 8
- select the amp setup script from the desktop on the computer
- load normalized dilutions and samples according to the amp worksheet
- all sample loading must be witness by another analyst
2
Q
explain the process of amplification with the qiagility - post-run
A
- check the bottom of the plate for volume differences
- place strip caps on the plate
- centrifuge the plate and start the thermocycler run
- re-cap and save the suspension buffer
- re-cap the extracts and store them; dilutions may be discarded
- discard all consumables except tips
3
Q
explain setting up capillary electrophoresis with the qiagility - post-run
A
- place septa on the plate and centrifuge
- sandwich the CE plate between the blue plate base and the white plate retainer and start the CE run
- wipe down the qiagility with ethanol and UV after
4
Q
what is the liquid level detection system
A
- for the qiagility to measure how much liquid is in each tube
- centrifuge all samples for 30 seconds to minimize errors with the LLD
- sample loading must be witnessed by another analyst
5
Q
what are some liquid level detection system errors and how are they solved
A
- the qiagility is trained to pipette all volumes required by LSPCL procedures
- the most common robot errors are with the LLD
- if there is an error “send” the robot arm back to its starting point and click “retry operation”
- if retrying does not solve the error, visually inspect the samples
- LLD errors are most commonly seen during amp setup, with dilution volumes below 15 uL
6
Q
explain the process of loading the qiagility for quant
A
- make powerquant master mix
- aliquot standard one from the powerquant kit into the first micro PCR tube
- fill the NTC tube with powerquant dilution buffer
7
Q
what are some pros and cons of the qiagility
A
- pros
- automation
- precision
- removes human error from pipetting
- cons
- care must be given during loading
- all samples are open at once
8
Q
what is the qiagility instrument
A
- a pipetting robot that performs fully automated PCR setups
- used to setup quant, amp, and CE plates
- all tubes must be uncapped
9
Q
what are the qiagility blocks for amp
A
- in M1, a 1.5 mL tube with suspension buffer goes in spot A
- an elution tube with master mix goes in spot B
10
Q
what are the qiagility blocks for post-amp/ce
A
- there are four tip racks instead of two
- A1, A2, B1, B2
- in R1, the Fusion 6C ladder goes in spot A
- in M1, a blue master mix tube goes in spot B
- blocks C1 and B2 are for 96-well plates
- the amp plate goes in block C1 (remove strip caps) and the empty CE plate goes in spot C2
11
Q
what are the qiagility blocks for quant
A
- A1 and A2 are for tips
- Blocks B1, B2, and C1 are for sample tubes
- block C2 is for a 96-well plate
- in M1, a 1.5 mL master mix tube goes in spot A
- in R1, the standard dilutions go in spots A1-A4
- the NTC goes in spot B1
- in R1, the dilution buffer goes in spot A
12
Q
list some qiagility components
A
- qiagility instrument, qiagility software, and a computer
- have been optimized to work together to provide accuracy, precision and speed
- best for large plates
13
Q
explain qiagility decontamination
A
- remove all reagents and consumables
- wipe down surfaces with ethanol
- don’t remove tips or wipe down the qiagility hood
14
Q
what are some qiagility parts
A
- hood
- hepa filter
- feet
- uv lamp
- tip chute
- robotic arm (pipetting head)
- master mix block
- reagent block
- tips, sample, and reaction blocks
- tip ejector
15
Q
how does the qiagility instrument perform
A
- uses a single-channel pipette to automate PCR/post-PCR setup
- takes 30 min for a full plate
- senses liquid levels, facilitated by conductive tips, and enables high-precision pipetting
- follows scripts to complete different procedures
16
Q
explain quantification with the qiagility - post-run
A
- always visually inspect the bottom of plates for volume differences
- place a clear seal on the powerquant plate, centrifuge, and start quantstudio 5 run
- cap all samples with new caps
- standards may be saved, but mark for training only
17
Q
what are the different types of qiagility scripts for quant
A
- machine-made standards
- the instrument will make the standard dilution series
- pre-made standards
- using standard dilutions that have already been made by another analyst
- good for seven days
18
Q
how do we uv the qiagility
A
- the UV must be run before and after every use of the qiagility
- takes 30 minutes to complete
- click the light bulb icon at the top of the screen, set the timer to 30 minutes, click “close software when UV finishes”, then click start
- do not click “enable lamp”
- the UV won’t shut off automatically
