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| Status |
Public on May 12, 2021 |
| Title |
RNA-CASing |
| Organism |
Escherichia coli |
| Experiment type |
Other
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| Summary |
The specificity of the RNA-CASing process was analysed by Next-Generation Sequencing. Therfor small RNAs were isolated from purified proteins of Escherichia coli and subjected to Illumina sequencing or nanopore sequencing.
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| Overall design |
cDNA libraries were created with the NEBNext® Multiplex Small RNA Library Pret Set for Illumunia according to the manufacturer’s instructions. 100 ng of input RNA was used and amplified cDNA libraries were separated by native PAGE for size selection of 120-250 bp. E. coli small RNA isolates were then subjected to Illumina. Extracted RNA was treated with E. coli Poly(A) Polymerase (NEB) for 30 min 37°C to add a poly(A) tail and then processed for Nanopore sequencing with the Direct RNA Sequencing Kit according to the protocol provided by Oxford Nanopore Technologies.
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| Contributor(s) |
Randau L |
| Citation missing |
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| |
| Submission date |
May 11, 2021 |
| Last update date |
May 14, 2021 |
| Contact name |
Lennart Randau |
| E-mail(s) |
lennart.randau@staff.uni-marburg.de
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| Organization name |
Phillips-Universität Marburg
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| Department |
Biology
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| Lab |
Randau
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| Street address |
Karl-von-Frisch-Str. 4
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| City |
Marburg |
| State/province |
Hessen |
| ZIP/Postal code |
35043 |
| Country |
Germany |
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| Platforms (2) |
| GPL18133 |
Illumina HiSeq 2500 (Escherichia coli) |
| GPL30119 |
MinION (Escherichia coli) |
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| Samples (3) |
| GSM5290693 |
Repeat-tagged sfgfp Illumina Sequencing |
| GSM5290694 |
Repeat-tagged lacZ-α Illumina Sequencing |
| GSM5290695 |
Repeat-tagged sfgfp Nanopore Sequencing |
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| Relations |
| BioProject |
PRJNA729041 |
| SRA |
SRP319379 |
