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Library preparation may be done manually or on our Takara Bio Apollo 324 Robot or our Eppendorf epMotion 5075 robot depending on the type of prep and number of samples. Some libraries are priced per sample, others have a setup fee with an additional cost per sample. The robotic preps are limited to the number of libraries that are done at the same time and are priced accordingly (ie. 6, 8, 96) Please contact Mark for more details.
Quality: A better quality sample will give a better library. Nucleic acids should have absorbance ratio values A260/A280 of 1.8–2.0. Longer length genomic DNA is better and for some preps necessary. RNA quality should be checked with the bioanalyzer before library preps and is not included in the fee. RINs of 8-10 are best for most preps. If the RIN is less, talk to us about possible preps.
Quantity: See chart below for specifics for prep type. We appreciate more DNA or RNA if you have sufficient material for 2X or more, especially if you have determined concentration by a spectrophotometer such as nanodrop rather than a fluorometric method such as Qubit. We will double check the concentration with the Qubit fluorometer before starting. You must supply enough extra DNA or RNA beyond the required amount for the prep. An additional 2-5 mL is recommended. This is included in the library prep fee.
PCR Cycles: We will use a recommended default value for the number of PCR cycles in the protocol’s amplification step. Notify us if you would like to change this number.
Size Determination: After library prep is complete, the size distribution must be checked with the bioanalyzer HS-DNA chip or TapeStation HS DNA tape. This is not included in the prep fee.
Size Selection: Some preps will need additional size selection. We will notify you if that is the case.
Library Quantification: We recommend the library prep concentration to be determined by qPCR before sequencing for many of our preps. This is an extra fee.
Pooling: Library pooling is included in the library prep fees. Let us know how you would like your libraries pooled and in what ratios before we start the prep. Selection of barcodes for the pool is important to plan before library prep is started.
|Library Prep||Minimum concentration (ng/mL)*||Minimum Volume for prep and QC (mL) !||Quantity DNA or RNA needed for prep (ng)||Insert or PCR amplicon size bp (w/o adapters)|
|PrepX DNA||1.5||105||150||220, 320, 520, 870|
|TruSeq DNA Nano 350 bp or 550 bp insert||1.5 or 2.5||105||150 or 250||350, 550|
|TruSeq DNA PCR-Free 350 bp or 550 bp insert||105 or 205||105||1050 or 2050||350, 550|
|Nextera XT DNA||0.2||7||1||200-2000 (835)|
|Illumina Nextera Flex DNA - Large Genome||3.5-17||35||100-500||300-350|
|Illumina Nextera Flex DNA - Small Genome||0.05-17||35||1-500||300-350|
|Nextera Mate Pair Gel-Free||14||80||1000||2-15kb (2.5-4kb)|
|Nextera Mate Pair Gel-Plus||13||312||4000||Gel size selection|
|TruSeq ChIP||0.17||65||10||150-200 (custom)|
|PrepX Robotic PolyA + Stranded RNA||20||55||1000||100-200|
|PrepX Robotic RiboZero + Stranded RNA||3.6-180||32||1000-5000||100-200|
|Lexogen QuantSeq 3' mRNA-Seq FWD||2-400||8||10-2000 (500)||100-200|
|TruSeq Small RNA||200||7||1000||22-30|
|16s amplicon PCR library||2.5||7||12.5||460|
|Ask us about methylation, Hi-C, other preps of interest|
*It is appreciated if you can supply a higher concentration and larger volume than the minimum recommendation. If you have read your concentration with a spectrophotometric method instead of a fluorometric method, you may need to supply much more than the minimum concentration to have enough for a prep.
! You must supply enough extra DNA or RNA beyond the required amount for the prep for QC purposes. An additional 2-5 mL is recommended.