Table2.
Oligonucleotides used for HT-5Pseq library preparation
| Primer name | Purpose | Sequence (5′-3′) |
|---|---|---|
| RNA_rP5_RND | To add UMI and common sequence to the RNAs (step 11) | rCrUrUrUrCrCrCrUrArCrArCrGrArCrGrCrUrCrUrUrCrCrGrArUrCrU rNrNrNrNrNrNrNrN |
| 5Pseq-dT | To do reverse transcription by using oligo-dT and add common sequence to the libraries (step 17) | GTGACTGGAGTTCAGACGTGTGCTC TTCCGATCT TTTTTTTTTT |
| 5Pseq-RT | To do reverse transcription by using random hexamer and add common sequence to the libraries (step 17) | GTGACTGGAGTTCAGACGTGTGCTC TTCCGATCT NNNNNN |
| Illumina compatible PE1.0 | To generate library by PCR (step 37) | ATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACG ACGCTCTTCCGATC∗T |
| PE2_MPX_01 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_02 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATACATCGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_03 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGCCTAAGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_04 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTGGTCAGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_05 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCACTGTGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_06 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATATTGGCGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_07 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGATCTGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_08 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTCAAGTGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_09 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCTGATCGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_10 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATAAGCTAGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_11 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGTAGCCGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_12 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_13 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTTGACTGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_14 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGGAACTGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_15 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTGACATGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_16 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGGACGGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_17 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCTCTACGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_18 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGCGGACGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_19 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTTTCACGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_20 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGGCCACGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_21 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCGAAACGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_22 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCGTACGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_23 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCCACTCGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_24 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGCTACCGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_25 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATATCAGTGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_26 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATGCTCATGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_27 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATAGGAATGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_28 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCTTTTGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_29 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATTAGTTGGTGACTGGAGT TCAGACGTGTGCTCTTCCGATC∗T |
| PE2_MPX_30 | To add multiplex barcode by PCR (step 37) | CAAGCAGAAGACGGCATACGAGATCCGGTGGTGACTGGAG TTCAGACGTGTGCTCTTCCGATC∗T |
∗
refers to S-linkage between the two bases r refers to a RNA base. Barcodes are identified in bold black. Sequences are represented in 5′ to 3′orientation. # Oligonucleotide sequences © 2006–2021 Illumina, Inc. All rights reserved.