New England Biolabs, E1050L, Exo-CIP™ Rapid PCR Cleanup Kit

Related Categories PCR & DNA Cleanup,, Nucleic Acid Purification Applications PCR & Reaction Cleanup,, DNA Amplification, PCR & qPCR FAQ Q: Do I have to purify my PCR amplicon after cleanup? A: No, PCR amplicons treated with Exo-CIP are immediately ready for downstream applications without any need for further processing. Q: How should I store my treated samples after the enzymatic cleanup reaction? A: We recommend storing treated samples at -20°C to ensure the PCR polymerase remains inactive. Q: Why are my sequence reads less than 600 bases even when the electropherogram looks great? A: This could be the result of submitting too much sample for sequencing and overloading the BigDye™ terminator chemistry. Make sure the submitted sample is between 15-200 fmol in a 3 µl volume. This corresponds to the following mass amounts: Size of PCR amplicon ng of DNA (in 3 ul sample) 100 bp 1 - 12 500 bp 5 - 60 1000 bp 10 - 120 3000 bp 30 - 360 5000 bp 50 - 600 Q: Why do I see two peaks (one large and one small) overlapping in one position of the electropherogram? A: This phenomenon may be explained by one of the two following possibilities. The first is more than one size of PCR product was amplified. If the amplification does not produce a homogeneous product and more than one species is amplified with sequence similarity to another, both can be sequenced with the added sequencing primers. Another explanation may be that unprocessed sample remained on the side of the reaction tube, leading to aberrant reads. Please be sure to vortex and quickly spin the reaction tube after treatment with the Exo-CIP reagents. Q: Is my master mix / PCR buffer compatible with the Exo-CIP Rapid PCR Cleanup Kit? A: No buffer change or additives are necessary. The Exo-CIP Rapid PCR Cleanup Kit has been tested for compatibility with the following master mixes and buffers. In addition, the routine buffers provided with typical Taq-based products from numerous vendors are also expected to be compatible with this product. If interference is observed (due to the presence of PCR enhancers, etc.), please dilute the final PCR product volume with an equal volume of nuclease–free water, prior to cleanup with Exo-CIP. Q5® High-Fidelity DNA Polymerase Q5® Hot Start High-Fidelity DNA Polymerase Q5® High-Fidelity 2X Master Mix Q5® Hot Start High-Fidelity 2X Master Mix NEBNext® Q5® Hot Start HiFi PCR Master Mix NEBNext® Ultra™ II Q5® Master Mix Phusion® Hot Start Flex 2X Master Mix OneTaq® Hot Start Quick-Load® 2X Master Mix with Standard Buffer OneTaq® Hot Start Quick-Load® 2X Master Mix with GC Buffer Taq 2X Master Mix LongAmp® Taq 2X Master Mix Taq DNA Polymerase with Standard Taq buffer Taq DNA Polymerase with Thermopol® buffer Phire™ Hot Start II DNA Polymerase Phire™ Hot Start II DNA Polymerase w/ DMSO Platinum™ SuperFi™ DNA Polymerase Phire™ Plant Direct PCR kit Kapa™ HiFi PCR Fidelity buffer Kapa™ HiFi PCR GC buffer Kapa™ HiFi HotStart Ready Mix PrimeSTAR® GXL DNA Polymerase Terra™ PCR Direct