... | ... | @@ -143,7 +143,24 @@ ORF prediction was done on the PacBio MCF-7 sequences using [ANGEL](http://www.g |
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---
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## **RNA-FISH and Sanger Validations** <br>
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To be completed ... <br>
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The PCR for Sanger sequence validation was performed using the primers (listed in the table below) using the 2x Phusion High-Fidelity PCR master mix with HF buffer (NEB). Briefly, the pcr ran for 30 cycles with 1 min elongation at 72C. The PCR products were purified using Ampure XP beads following the guidelines of the manufacturer. The sizing of the amplicons was checked using Agilent’s Labonachip system. The Sanger sequencing of the products was performed by the LGTC and the sequences were analyzed using Sequence Scanner Sofware 2 (Applied Biosystems, CA USA).<br>
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|Gene |Fragment |5' Primer |3' Primer |
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|:---|:---|:---|:---|
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|C1QTNF6 |A|GCTCCACCATAGGGATCTCA|CAGAGGGAGAACGGAACAAA|
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|C1QTNF6 |B|GCTCCACCATAGGGATCTCA|TTAGTATTGCAGACATGGGCC|
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|C1QTNF6 |C|GGCCCATGTCTGCAATACTAA|CAGAGGGAGAACGGAACAAA|
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|C1QTNF6 |D|GGCCCATGTCTGCAATACTAA|CCCCCTTCTTGAGCTATTTACA|
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|CALU |A|ACGGGTTTGTCACTGTGGAT|CCATTTTAAACCTCCGCTCA|
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|CALU |B|CAGAAACGTGACTTATGGCA|CCATTTTAAACCTCCGCTCA|
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|CALU |C|ATGGACCTGCGACAGTTTCT|CCATTTTAAACCTCCGCTCA|
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|CALU |D|ACGGGTTTGTCACTGTGGAT|TGCCATAAGTCACGTTTCTG|
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|ALDOA|A|GCTTCGAGATCAAGCTCCGA|ATTCAAGGAGAGAACGCGGG|
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|ALDOA|B|GCCTCCGTCTGGATTTCCAA|ATTCAAGGAGAGAACGCGGG|
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|ALDOA|C|GCTTCGAGATCAAGCTCCGA|GCGATGTCAGACAGCTCCTT|
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|ALDOA|D|CCCGCGTTCTCTCCTTGAAT|CAACACCGCCCTTGGATTTG|
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|RELA|A|ATGGCTCGTCTGTAGTGCAC|GCCTGGTCCCGTGAAATACA|
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|RELA|B|CTGTTCCCCCTCATCTTCCC|GATCTTGAGCTCGGCAGTGT|
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<p align="right"> [`TOP`](#full-length-mrna-sequencing-uncovers-a-widespread-coupling-between-transcription-and-mrna-processing)</p><br>
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... | ... | @@ -193,9 +210,14 @@ Leiden University Medical Center <br> |
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Department of Human Genetics <br>
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Leiden, 2300 RC, The Netherlands <br>
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<br>
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**Raymund Yin** <br>
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LGC Biosearch Technologies <br>
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Petaluma, CA 94954-6904, USA <br>
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<br>
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**Hans Johansson** <br>
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LGC Bioresearch Technologies <br>
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LGC Biosearch Technologies <br>
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Petaluma, CA 94954-6904, USA <br>
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<br>
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