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View additional product information for Ion AmpliSeq™ Library Kit Plus - FAQs (A38875, 4488990, A35907)
9 product FAQs found
When handling barcode adapters, be careful to avoid cross-contamination by changing gloves frequently and by carefully removing the plate seal.
- Pulse vortex the IonCode Barcode Adapters plate several times.
- Centrifuge the IonCode Barcode Adapters plate for 1 minute at 1000 x g to collect contents.
- Carefully remove, then discard the plate seal.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
The IonCode Barcode Adapters 1-96 Kit consists of 1 x 96-well plate, with 20 µL of each barcode per well. The plate consists of 960 reactions in total with 10 reactions for each of the 96 barcodes.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
The IonCode Barcode Adapters 1-96 Kit consists of 1 x 96-well plate, with 20 µL of each barcode per well. The barcodes are pre-diluted, ready-to-use. The 96-well plate format allows multichannel pipetting for more efficient work.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
The IonCode Barcode Adapters 1-96 Kit is shipped on dry ice and we recommend storing at -30 degrees C to -5 degrees C in an upright position.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
Here are possible causes and solutions:
- The sample DNA or RNA may have been misquantified. We recommend requantifying the DNA or RNA. See the user guide for your library preparation method.
- More than 100 ng of sample DNA or RNA may have been used. We recommend decreasing the number of amplification cycles by adding less DNA or RNA.
- If you are using the Agilent 2100 Bioanalyzer instrument, the markers may have been misassigned. We recommend ensuring that the markers are correctly assigned.
- The library may have been contaminated by high molecular weight DNA. We recommend one of the following:
a) Removing less supernatant in the first-round (0.5X) purification and to ensure that the bead pellet is not disturbed.
b) Increasing the AMPur XP Reagent volume from 25 µL (0.5X) to 35 µL (0.7X) in the first-round purification.
- Inserts may have concatemerized during the ligation steps. We recommend reducing nucleic acid input, requantifying samples with a Qubit Fluorometer, or reducing the target amplification cycle number.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
- The sample DNA or RNA may have been misquantified. We recommend requantifying the DNA or RNA. See the user guide for your library preparation method.
- Target amplification may have been inhibited by residual ethanol in the sample DNA or RNA. We recommend incubating the tube uncapped in a hood for 1 hr or using a Speed-vac for 5 mins at room temperature to remove excess ethanol.
- Library amplification may have been inhibited by residual ethanol from the AMPure purification. We recommend carefully removing all drops of ethanol before library amplification, then centrifuging the plate, if needed.
- The sample DNA or RNA may have been of low quality. We recommend adding more DNA or RNA or increasing the number of amplification cycles.
- PCR, digestion, or ligation may have been inefficient. We recommend ensuring proper dispensing and mixing of viscous reagents at each step.
- The AMPure XP Beads may have been over-dried. We recommend drying the beads for 5 mins. Do not over dry the beads.
- The AMPure XP Beads may have inhibited library amplification. We recommend transferring the library from the beads before amplification.
- The qPCR cycling time may have been too short. We recommend using standard qPCR cycling conditions instead of fast cycling for library designs >175 bp.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
We recommend storing at -30 to -10 degrees C except for Low TE which should be stored at 15 to 30 degrees C.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
This 384-reaction size kit is composed of four 96-reaction size kits, Cat. No. A35907. Each 96-reaction size kit contains the following components:
- 5X Ion AmpliSeq HiFi Mix (red cap), 480 µL
- FuPa Reagent (brown cap), 192 µL
- Switch Solution (yellow cap), 384 µL
- DNA Ligase (blue cap), 192 µL
- 25X Library Amp Primers (pink cap), 192 µL
- 1X Library Amp Mix (black cap), 4 x 1.2 mL
- Low TE, 2 x 6 mL
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
The kit is available in the following reaction sizes:
- 24 reactions (Cat. No. 4488990)
- 96 reactions (Cat. No. A35907)
- 384 reactions (Cat. No. A38875)
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.