Total RNA from tumor cell lines was isolated using RNeasy Protect Mini Kit (QIAGEN) according to the manufacturer’s instructions. cDNA preparation was performed according to the standard procedures using M-MLV Reverse Transcriptase (Promega) and oligo-deoxythymine primers/random hexamers. Polymerase chain reaction (PCR) was performed by applying the following TaqMan probes: Hs00367063_m1(PLXNB2), Hs00195591_m1(SNAI1), Hs03676575_s1(ID1), Hs00357821_g1(ID1), Hs00954037_g1(ID3), s01023894_m1(CDH1), and Hs00950344_m1(SNAI2). Alternatively, PCR was conducted with SYBR Green Master Mix (Life Technologies) and run in Applied Biosystems 7900HT Fast Real-Time PCR System, by applying the following primer pairs: glyceraldehyde-3-phosphate dehydrogenase [5′-GAAGGTGAAGGTCGGAGTC-3′ (forward) and 5′-GAAGATGGTGATGGGATTTC-3′ (reverse)], Snail [5′-GACTACCGCTGCTCCATTCCA-3′ (forward) and 5′-TCCTCTTCATCACTAATGGGGCTTT-3′ (reverse)], Slug [5′-AGATGCATATTCGGACCCAC-3′ (forward) and 5′-CCTCATGTTTGTGCAGGAGA-3′ (reverse)], Zeb1 [5′-AGCAGTGAAAGAGAAGGGAATGC-3′ (forward) and 5′-GGTCCTCTTCAGGTGCCTCAG-3′ (reverse)], Twist1 [5′-TGTCCGCGTCCCACTAGC-3′ (forward) and 5′-TGTCCATTTTCTCCTTCTCTGG-3′ (reverse)], h–E-cadherin [5′-GTCACCTTCAGCCATCCTGT-3′ (reverse) and 5′-GGGTTATTCCTCCCATCAGC-3′ (forward)], h-PlexinB1 [5′-CACTGAACCCCACACCTTTC-3′ (forward) and 5′-ATAGCCACCACCTCCTCCTT-3′ (reverse)], h-PlexinB2 [5′-CTTGACCTGGGAGATGGTGT-3′ (reverse) and 5′-CTGGGGGATGATGTGGAGTA-3′ (forward)], and h-Sema4C [5′-GACACCTCCTGGCACAACAC-3′ (forward) and 5′-CCACTTCTGGGCTTCCTCA-3′ (reverse)].

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.