Editing BioMicroCenter:RNA LIB (section)

== [https://www.neb.com/products/e6310-nebnext-rrna-depletion-kit-human-mouse-rat#Product%20Information NEB Ultra II Directional RNA with rRNA Depletion (H/M/R)] ==
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  !Service 
  !Standard RNA Library Prep
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  |SAMPLE INPUT || [[BioMicroCenter:RIN|Intact]] or [[BioMicroCenter:RIN|degraded]] total RNA
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  |RANGE OF INPUT || 5-100 ng
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  |INCLUDED || Initial QC by Fragment Analyzer <BR> Library preparation <BR> Illumina QC
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  |SEQUENCING RECOMMENDATIONS || All platforms
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  |INDEX AVAILABILITY || 112 [[BioMicroCenter:HTLR_TEST_HTLR_TEST#Unique_versus_Combinatorial_Dual_Indexing|Unique Dual Indexes]] <br> 192 [[BioMicroCenter:HTLR_TEST_HTLR_TEST#Unique_versus_Combinatorial_Dual_Indexing|Combinatorial Dual Indexes]]
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  |SUBMISSION || MIT - [https://mit.ilabsolutions.com/service_item/new/3381?spt_id=3863 ilabs] <BR> External - [[BioMicroCenter:Forms|form]]
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  |UNIT || Per sample
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NEBNext Ultra II Directional RNA preparation with ribosomal RNA (rRNA) depletion is used to deplete rRNA by enzymatic degradation using single-stranded DNA probes that target rRNAs, leaving all other RNA species present and available for library preparation. This depletion method is agnostic to input sample quality and is the go-to method if samples don't meet quality requirements for poly(A) selection. This method is suited for inputs ranging from 5ng to 100ng (recommended input of > 50ng). Due to probe design used in the depletion, this method is currently only available for Human/Mouse/Rat samples. We also offer a [[BioMicroCenter:RNA_HTL|high-throughput protocol]].
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==[https://sequencing.roche.com/en-us/products-solutions/by-category/library-preparation/rna-library-preparation/kapa-mrna-hyperprep-kits.html Kapa mRNA Hyperprep] ==
{|
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  !Service 
  !Standard RNA Library Prep
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  |INPUT || Clean eukaryotic total RNA <BR> RIN 9+ <BR> >10ng/uL <BR> >10uL
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  |INCLUDED || Initial QC by Fragment Analyzer <BR> Library preparation <BR> Illumina QC
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  |INDEX AVAILABILITY || 96 UDI
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  |SEQUENCING RECOMMENDATIONS || All platforms
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  |SUBMISSION || MIT - [https://mit.ilabsolutions.com/service_item/new/3381?spt_id=3863 ilabs] <BR> External - [[BioMicroCenter:Forms|form]]
  |-
  |UNIT || Per sample
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This workflow is very similar to Illumina's TruSeq chemistry at lower cost and is streamlined for automation. This chemistry uses polyT beads to isolate the mRNA from the rRNA and tRNA. The use of these beads requires that the RNA be of very high quality or only the 3' end of transcripts will be isolated. Purified mRNA is then fragmented with metal and random priming is used to convert the sample to cDNA. Once double-stranded cDNA is generated, LMPCR is performed to create the indexed Illumina library. The BioMicro Center offers mRNA HyperPrep as a single sample reaction or in [[BioMicroCenter:RNA_HTL|batches of 24 done on the TecanEvo 150s.]] 
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== [https://sequencing.roche.com/en-us/products-solutions/by-category/library-preparation/rna-library-preparation/kapa-rna-hyperprep-kit-with-riboerasehmr.html Kapa RiboErase] & [https://www.illumina.com/products/by-type/accessory-products/ribo-zero-plus-rrna-depletion.html RiboZero] ==
{|
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 {| class="wikitable" border=1
  !Service 
  !rRNA depletion based RNA Library Prep
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  |INPUT || Clean total RNA <BR> DV200>0.2 <BR> 
Human/Mouse/Rat
* >10ng/uL 
* >10uL
All others
* > 200ng/uL
* > 10uL
  |-
  |INCLUDED || Initial QC by Fragment Analyzer <BR> Library preparation <BR> Illumina QC
  |-
  |SUBMISSION || MIT - [https://mit.ilabsolutions.com/service_item/new/3381?spt_id=3863 ilabs] <BR> External - [[BioMicroCenter:Forms|form]]
  |-
  |UNIT || Per sample
 |}
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For samples with degraded RNA or samples where you are interested in looking at non-polyA RNAs, the BioMicro Center utilizes the Kapa RNA RiboErase and Illumina RiboZero for Human/Mouse/Rat samples. RiboErase uses RNAseH to degrade rRNAs while RiboZero uses magnetic beads coupled to rRNA sequences to remove these sequences from the solution. The remaining mRNA fragments can then be converted in to cDNA and prepared using the NEB Ultra II Directional RNA or Kapa mRNA Hyperprep kit to produce the Illumina library.<BR><BR>
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