---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2010-01-14 19:23:17+00:00
layout: post
slug: solid-bead-titration-herring-fragmented-cdna-libraries-2lhkod09-4lhtog09-6lhpws09
title: 'SOLiD Bead Titration - Herring fragmented cDNA libraries: 2LHKOD09, 4LHTOG09,
  6LHPWS09'
categories:
  - 2010
  - Miscellaneous
tags:
  - 2LHKOD09
  - 4LHTOG09
  - 6LHPWS09
  - bead counts
  - cDNA
  - library prep
  - Pacific herring
  - SOLiD
  - SOLiD libraries
  - templated bead prep
---

Continued with templated bead prep from ePCRs for these libraries. Samples were processed according to the ABI "Templated Bead Preparation Guide" following the "full-scale" protocol.

To see explanations of the various calculations below, see the "SOLiD Bead Titration - Herring fragmented cDNA library 3LHSITK09 (CONTINUED from ePCR yesterday)" from 20100107.

2LHKOD09:

Initial counts: 109, 129, 112, 115 ---- Avg. = 116.25 beads/square

Beads: 116.25 x 10 x 25 x 200 = 5.8125x10^6 beads/uL x 200uL = 1.1625x10^9 beads

Templated beads counts: 205, 199, 197, 210 ----- Avg. = 210 beads/square

**Templated beads**: 210 x 10 x 25 x 10 = 5.06875x10^5 beads/uL x 400uL = **2.0275x10^8 beads**

Efficiency: 17.44%

4LHTOG09:

Initial counts: 124, 124, 117, 104 ----- Avg. = 117.25 beads/sqaure

Beads: 117.25 x 10 x 25 x 200 = 5.8625x10^6 beads/uL x 200uL = 1.1725x10^9 beads

Templated beads counts: 139, 135, 145, 140 ---- Avg. = 139.75 beads/square

**Templated beads**: 139.75 x 10 x 25 x 10 = 3.49375x10^5 beads/uL x 400uL = **1.3975x10^8 beads**

Efficiency: 11.92%

6LHPWS09:

Initial counts: 135, 106, 123, 124 ---- Avg. = 122 beads/square

Beads: 122 x 10 x 25 x 200 = 6.1x10^6 beads/uL x 200uL = 1.22x10^9 beads

Templated beads counts: 141, 171, 164, 170 ----- Avg. = 161.5 beads/square

**Templated beads**: 161.5 x 10 x 25 x 10 = 4.0375x10^5 beads/uL x 400uL = **1.615x10^8 beads**

Efficiency: 13.24%

All beads were stored @ 4C until ready for bead deposition and work flow analysis run.

Results:

Rhonda Morales (from Ginger's lab who is responsible for running/maintaining the SOLiD at the CEG) says the numbers on all samples look perfect! Will proceed to work flow analysis once Jesse's samples are ready (ETA of Jan. 27th, 2010).

Here are [reagent lot numbers for the Bead Titration](https://eagle.fish.washington.edu/Arabidopsis/Notebook%20Workup%20Files/20100116-01.jpg).
