---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2015-12-22 22:51:34+00:00
layout: post
slug: illumina-methylation-library-quantification-bs-seq-olyc-gigas-libraries
title: Illumina Methylation Library Quantification - BS-seq Oly/C.gigas Libraries
categories:
  - 2015
  - BS-seq Libraries for Sequencing at Genewiz
  - Olympia oyster reciprocal transplant
tags:
  - 1NF11
  - 1NF15
  - 1NF16
  - 1NF17
  - 2NF5
  - 2NF6
  - 2NF7
  - 2NF8
  - BS-seq
  - Crassostrea gigas
  - DNA Quantification
  - Katherine Silliman
  - Katie Lotterhos
  - M2
  - M3
  - NF2_6
  - NF_18
  - olympia oyster
  - Ostrea lurida
  - Pacific oyster
  - Qubit 3.0
  - Qubit dsDNA HS
  - TruSeq DNA Methylation Library
---

Re-quantified [the libraries that were completed yesterday](https://robertslab.github.io/sams-notebook/posts/2015/2015-12-22-illumina-methylation-library-construction-olyc-gigas-bisulfite-treated-dna/) using the Qubit3.0 dsDNA HS (high sensitivity) assay because the library concentrations were too low for the normal broad range kit.

Results:

Qubit Quants and Library Normalization Calcs: [20151222_qubit_illumina_methylation_libraries](https://docs.google.com/spreadsheets/d/1bfIDqNPOxnShlP3Esl0D0pDOSXc5Yn6Ar4RQVS3RJLo/edit?usp=sharing)

<table cellpadding="0" cellspacing="0" border="0" dir="ltr" > 
<tbody >
<tr >

<td data-sheets-value="[null,2,"Test Name"]" >**SAMPLE**
</td>

<td data-sheets-value="[null,2,"Original sample conc."]" >**CONCENTRATION (ng/μL)**
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"1NF11"]" >1NF11
</td>

<td data-sheets-value="[null,3,null,2.42]" >2.42
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"1NF15"]" >1NF15
</td>

<td data-sheets-value="[null,3,null,1.88]" >1.88
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"1NF16"]" >1NF16
</td>

<td data-sheets-value="[null,3,null,2.74]" >2.74
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"1NF17"]" >1NF17
</td>

<td data-sheets-value="[null,3,null,2.54]" >2.54
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"2NF5"]" >2NF5
</td>

<td data-sheets-value="[null,3,null,2.72]" >2.72
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"2NF6"]" >2NF6
</td>

<td data-sheets-value="[null,3,null,2.44]" >2.44
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"2NF7"]" >2NF7
</td>

<td data-sheets-value="[null,3,null,2.38]" >2.38
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"2NF8"]" >2NF8
</td>

<td data-sheets-value="[null,3,null,1.88]" >1.88
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"M2"]" >M2
</td>

<td data-sheets-value="[null,3,null,2.18]" >2.18
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"M3"]" >M3
</td>

<td data-sheets-value="[null,3,null,2.56]" >2.56
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"NF2_6"]" >NF2_6
</td>

<td data-sheets-value="[null,3,null,2.5]" >2.5
</td>
</tr>
<tr >

<td data-sheets-value="[null,2,"NF_18"]" >NF_18
</td>

<td data-sheets-value="[null,3,null,2.66]" >2.66
</td>
</tr>
</tbody>
</table>



Things look pretty good. The [TruSeq DNA Methylation Library Kit (Illumina)(https://github.com/sr320/LabDocs/blob/master/protocols/Commercial_Protocols/Illumina_truseq-dna-methylation-library-prep-guide-15066014-a.pdf) suggests that the libraries produced should end up with concentrations >3ng/μL, but we have plenty of DNA here to make a pool for running on the HiSeq2500.