We analysed genomic DNA after multiplex ligation-dependent probe amplification (MLPA) using the SALSA MLPA kit (MRC-Holland, Amsterdam, Netherlands), according to the manufacturer’s instructions. The PCR fragments were separated by capillary electrophoresis using Life Technologies 3500 Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA, USA). We analysed the MLPA data using Coffalyser.Net v.140721.1958 (MRC-Holland, Amsterdam, Netherlands). Probe ratios between 0.75 and 1.3 were considered within the normal range. A probe ratio of < 0.75 or > 1.3 indicated deletion or gain, respectively. A probe ratio of < 0.25 or > 1.8 indicated biallelic deletion or amplification. We used the SALSA MLPA P335 ALL-IKZF1 probemix to detect alterations of EBF1, IKZF1, CDKN2A, CDKN2B, PAX5, ETV6, RB1 and BTG1. We used SALSA MLPA P327 iAMP21-ERG probemix to detect alterations in ERG and the intrachromosomal amplification of chromosome 21. We used SALSA MLPA P329 CRLF2-CSF2RA-IL3RA probemix to detect P2RY8-CRLF2 (PAR1 deletion)38 (link).
Life technologies 3 500 genetic analyzer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Life Technologies 3,500 Genetic Analyzer is a capillary electrophoresis system designed for DNA sequencing and fragment analysis. It features a 96-capillary array, automated sample handling, and data analysis software. The core function of the instrument is to separate and detect fluorescently-labeled DNA fragments.
Automatically generated - may contain errors
4 protocols using life technologies 3 500 genetic analyzer
1
Multiplex Ligation-Dependent Probe Amplification for Genetic Alteration Detection
2
MLPA for Genetic Aberrations in Leukemia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA was analyzed using the SALSA MLPA kit (MRC-Holland, Amsterdam, the Netherlands), according to manufacturer’s instructions. The PCR fragments were separated by capillary electrophoresis on a Life Technologies 3,500 Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA, USA). MLPA data were analyzed using Coffalyser.Net v.140721.1958 (MRC-Holland, Amsterdam, The Netherlands). Probe ratio between 0.75 and 1.3 were considered to be within the normal range. Probe ratio below 0.75 or above 1.3 indicated deletion or gain, respectively. Probe ratio below 0.25 or above 1.8 indicated biallelic deletion or amplification. SALSA MLPA P335 ALL-IKZF1 probemix was used for detection of alterations of EBF1, IKZF1, CDKN2A, CDKN2B, PAX5, ETV6, RB1 and BTG1 genes. SALSA MLPA P327 iAMP21-ERG probemix was used for detecting alterations of ERG gene and iAMP21. SALSA MLPA P329 CRLF2-CSF2RA-IL3RA probemix was used for detecting P2RY8-CRLF2 (PAR1 deletion).
3
TP53 Structural Alterations Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect structural alterations of TP53, multiplex ligation‐dependent probe amplification (MLPA) reactions were carried out according to the manufacturer’s instructions. The MRC‐Holland SALSA MLPA probe mix P056‐C1 kit (MRC Holland) was used. PCR fragments were separated by capillary electrophoresis on a Life Technologies 3500 genetic analyzer (Thermo Fisher Scientific). MLPA data were analyzed using Coffalyser.Net v.140721.1958 (MRC Holland). Relative copy numbers were determined after normalization of peaks against controls. Values between 0.75 and 1.3 were considered within the normal range. Values below 0.75 or above 1.3 implied a deletion or gain, respectively. Values below 0.25 indicated biallelic deletion.
4
MLPA Analysis of Leukemia Genes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA was analyzed by the SALSA multiplex ligation-dependent probe amplification (MLPA) kit (P335-A4) (MRC-Holland, Amsterdam, the Netherlands) according to suggestions of the manufacturer. This kit includes probes for IKZF1 (7p12.2, exons 1 to 8), CDKN2A (9p21.3, exons 2 and 4), CDKN2B (9p21.3, exon 2), PAX5 (9p13.2, exons 1, 2, 5, 6, 8, and 10), EBF1 (5q33.3, exons 1, 10, 14, and 16), ETV6 (12p13.2, exons 1, 2, 3, 5, and 8), BTG1(12.q22, exon 1 and 2), RB1 (13q14.2, exons 6, 14, 19, 24, and 29), SHOX (Xp22.33), CSF2RA (Xp22.33, exon 16), IL3RA (Xp22.33, exon 1), and CRLF2 (Xp22.33, exon 4). The PCR fragments were separated by capillary electrophoresis on a Life Technologies 3,500 Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA). MLPA data were analyzed with an ABI 3,730 capillary sequencer (Applied Biosystems, Foster City, CA). The relative copy number was determined after the normalization of peaks against controls. Values between 0.75 and 1.3 were considered within the normal range. Values below 0.75 or above 1.3 indicated deletion or gain, respectively. Values below 0.25 indicated biallelic deletion28 (link),29 (link).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!