Workflow
The qPCR Extraction Control is added to the lysis buffer along with the test sample before extraction to monitor poor extraction yield, PCR inhibition, incorrect pipetting or cycling parameters.
Control mix (includes primer and probes for control) is added to the reaction mixture
LYSIS & EXTRACTION
qPCR Extraction Control
ASSAY SET-UP
qPCR RESULTS
Possible Result Target
Internal Control DNA Interpretation
Test sample
1
+
+
Target(s) and internal control DNA detected.
Target(s) not detected, internal control DNA detected, indicates a sucessful extraction and qPCR reaction. Invalid result: Target(s) and internal control DNA not detected, repeat test.
2
-
+
qPCR extraction control starts monitoring from lysis and extraction
3
-
-
4
+
-
Invalid result: Internal control not detected, repeat test.
Product Highlights
qPCR Extraction Control Performs Identical To Spiked Internal Control DNA In Monitoring Inhibition
A) Spiked Internal Control DNA (green channel) and B) qPCR Extraction Control (red channel) were amplified from HEK293 cells. A standard column based genomic DNA extraction kit was used, with the lysis buffer or binding buffer being substituted with PBS to simulate inefficient extraction. Conditions were 10 min at 95 °C 10 s, 60 °C 30 s. Complete lysis step (red) and the pattern on inhibition for no lysis (orange) and no binding buffer (green) are the same across both types of controls.
A
B
Suitable For Multiplex Or Singleplex Reactions
A
B
C
A) A fragment of the beta 2 microglobulin (ß2MG) gene and B) A fragment of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene were amplified from HEK293 cells (green channel) and C) the internal control sequence were amplified from the qPCR Exctraction Control. Conditions were 10 min at 95 °C followed by 50 cycles 95 °C 10 s, 60 °C 30 s. The results illustrate that there is no difference in Cts between singleplex and multiplex reactions.
USA E: info@meridianlifescience.com Toll free: +1 800 327 6299 UK E: info.uk@meridianlifescience.com Tel: +44 (0)20 8830 5300 Ordering information:
Germany E: info.de@meridianlifescience.com Tel: +49 (0)3371 60222 00 France E: info.fr@meridianlifescience.com Tel: +33 (0)1 42 56 04 40
美国迈迪安生命科学公司 电子邮件: vivian.li@meridianlifescience.com 电话: +86-159-1103-0750
Connect with us:
Australia E: info.au@meridianlifescience.com Tel: +61 (0)2 9209 4180
www.MeridianLifeScience.com
ISO 13485 Certified
10/19
Powered by FlippingBook