Originally posted by atcghelix
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I routinely shear a couple hundred ng of DNA with a Covaris then concentrate it down to 20ul with an Amicon Ultra-0.5 10k. From there I can scale down my Kapa Hyper Prep reagents to 0.4x of what they should be. Haven't seen any problems yet! -
If you are going to do lots of library preps, it might be worth to do a comparison yourself. Prepping two libraries from the same input DNA and using 1/2 reactions with one and full reaction with other and comparison of PCR yield without sequencing would give some indications on performance of 1/2 reaction. With TruSeq kit, 1/2 and 1/3 reactions works as good as full reactions. Generally most kits use excessive amount of reagents to ensure good performance even with poor laboratory practices and low quality input.Leave a comment:
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Stretching Kapa library preps
Hi Everyone,
Has anyone tried doing half reactions with the Kapa library prep kits using 1000ng of input DNA? I'm curious if the amount of reagents in a half-reaction is enough to ligate adapters to 1000ng of input DNA.
Thanks very much!
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