---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2010-05-26 16:39:19+00:00
layout: post
slug: solid-epcrtemplated-bead-prep-lake-trout-lean-library
title: SOLiD ePCR/Templated Bead Prep - Lake Trout Lean library
categories:
  - 2010
  - Miscellaneous
tags:
  - bead counts
  - ePCR
  - lake trout
  - Lean
  - library prep
  - SOLiD
  - SOLiD libraries
  - templated bead prep
---

ePCR was performed following ABI's "full scale" protocol, using 1pM of SOLiD cDNA library.

Templated bead preparation was performed according to the "full scale" protocol.

Bead counts are calculated as follows:

Avg bead count x # hemacytometer squares x volume in hemacytometer (uL) x dilution factor = beads/uL x suspension volume (uL) = total beads

Initial Bead count: (1:200 dilution)

Lean: 126, 138, 122, 138 Avg. = 131

Lean: 131 x 25 x 10 x 200 = 6.55x10^6 beads/uL x 200uL = 1.31x10^9 beads

Templated Bead counts (1:10 dilution)

Lean: 165, 171, 186, 160 Avg. = 170.5

170.5 x 25 x 10 x 10 = 426250 beads/uL x 400uL = **1.705x10^8 beads**

Percent Recovery Templated Beads

Lean: (1.705x10^8 beads)/(1.31x10^9 beads) x 100 = **13.02% recovery**

Results: Yield is significantly higher than the previous preparation performed with this sample. The percent recovery falls into the desired range of 5-15%, so things look good there, too. Will contact Rhonda and get info regarding when this and the other 7 samples can go on a run.