Excellent ChIP-seq Preparation Kit

Carnegie Institution for Science
Embryology
Predoc fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Rubicon Genomics

Product Name:

ThruPLEX® DNA-seq Kit

Catalog Number:

R400429

Aim: study the epigenetic changes during mouse ES differentiation for different lineages. Specific goals: prepare ChIP-seq DNA library for histone H3K4Me3 modification for illumina high-seq 2000. Advantages: 1) less hands-on time 2) only one time Ampure beads clean after PCR amplification of library and minimal loss during preparation process 3) simple protocol and easy for new users who have no experience preparing ChIP-seq library 4) High DNA library yield

Experimental Design and Results Summary

Application

Mouse ES cell H3K4Me3 ChIP-seq library

Starting Material

300,000 ES cells

Protocol Overview

1) PurifyH3K4Me3 pulldown chromatin DNA ~100pg 2) Elute DNA into 10 ul and then follow exact template preparation and library synthesis steps from instruction. 3) Only use 10 PCR cycles for library amplification step (Kit suggests 14 cycles for 200pg DNA input). 4) Ampure beads purify ChIP-seq DNA library and elute in 30 ul. Take 1ul for bioanalyzer analysis.

Tips

The only inconvenient handling is small volume of enzyme and buffer mixture added into reaction during template preparation and synthesis steps.

Results Summary

1) Very good Library profile from bioanalyzer results. 2) Only takes me 3 hours to finish the whole process. I had to spend the whole day ~14h using illumina kit.

Additional Notes

None

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Summary

The Good

Less time and high quality

The Bad

None

The Bottom Line

I recommend this kit for your Library preparation for next-generation sequencing.

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