User guide

25
GlobalFiler
™
Express PCR Amplification Kit User Guide
Chapter2 Perform PCR
Swab substrates: prepare reactions
2
11. Amplify the samples in a Veriti
®
96-well Thermal Cycler or PCR System 9700
with the silver or gold-plated silver 96-well block as described in “Perform PCR”
on page 28.
IMPORTANT! This kit is not validated for use with the GeneAmp PCR System
9700 with the aluminium 96-well block. Use of this thermal cycling platform may
adversely affect performance of the this kit.
Swab substrates: prepare reactions
Sample prep
guidelines
• Detach each buccal swab head from the swab shaft before lysis.
• If using the heated lysis protocol, perform lysis in either of the following formats:
– 1.5 mL tubes with a heat block (VWR
®
Scientific Select dry heat block or
similar)
– 96-well deep-well plate (Part no. 4392904) with an oven and a metal plate
adaptor (Robbins Scientific
®
Model 400 Hybridization Incubator or similar,
Agilent
®
Benchtop Rack for 200 µl Tubes/V Bottom Plates (metal)
Part no. 410094 or similar)
IMPORTANT! Do not use a plastic plate adaptor.
• For optimum performance, lysis of a whole swab is recommended. To preserve
the sample, evaluate lysis of a half swab.
Prepare the
sample lysate
(room temperature
protocol)
1. Add 400 µL Prep-n-Go
™
Buffer (Part. no. 4471406) to 1.5 mL tubes or the
appropriate wells of a 96-well deep-well plate (Part no. 4392904).
2. Into each tube or well, put the entire head of each swab and let stand for
20 minutes at room temperature (20 to 25°C) to lyse the sample.
3. After 20 minutes, transfer the sample lysate out of the sample plate into tubes or
plates for storage, then discard the deep-well plate containing the swab heads.
Note: To minimize the risk of contamination, do not remove the swab heads from
the sample lysate plate before transferring the lysate.
4. Proceed to “Prepare the reactions” on page 21 or see “Store the sample lysate” on
page 27.
Prepare the
sample lysate (heat
protocol)
This protocol may improve the performance for challenging or aged samples.
1. Preheat the heat block to 90°C or the oven with metal plate adaptor to 99°C.
2. Add 400 µL Prep-n-Go
™
Buffer (for buccal swabs, Part. no. 4471406) to 1.5 mL
tubes or the appropriate wells of a 96-well deep-well plate (Part no. 43929040).
3. Into each tube or well, put the entire head of each swab. If you are using tubes,
cap the tubes. Let the tubes or plate stand for 20 minutes in the preheated heat
block or oven to lyse the sample.
4. After 20 minutes, remove the tubes or the deep-well plate from the heat block or
oven.