KAPA Single-Indexed Adapter Kits for Illumina plat

KAPA Single-Indexed Adapter Kits contain high-quality, ready-to-use adapters for Illumina library
sequencing applications. KAPA Single-Indexed Adapters are compatible with KAPA library prepar

KAPA Single-Indexed Adapters are available in two concentrations, namely 30 M and

well as fragment (insert) length and desired adapter:insert molar ratio (for DNA applications onl

- The most appropriate Adapter Stock Concentration for your application

- The factor by which that KAPA Single-Indexed Adapter stock should be diluted
- The number of libraries that can be prepared from the kit concentration and configuration (12
umina platforms
ers for Illumina library construction. Each adapter includes a single, 6-nt index (barcode) for multiplexed
th KAPA library preparation kits for DNA and RNA sequencing applications.

s, namely 30 M and 1.5 M. Based on your library construction kit and workflow, DNA or RNA input, as
r DNA applications only), this calculator is designed to return the following:

and configuration (12 or 24 adapters) you have selected

h adapter includes a single, 6-nt index (barcode) for multiplexed
and RNA sequencing applications.

n your library construction kit and workflow, DNA or RNA input, as

s designed to return the following:

u have selected
KAPA Single-Indexed Adapter Kit Calculator:

Instructions: Enter the appropriate values in the yellow and orange cells. Refer to the

Input into library construction 1

Mode size of fragmented DNA 200

Moles of insert DNA 0.01

Adapter:insert molar ratio 200.0

Moles of adapter needed per library 1.5

Volume of Adapter to use per ligation reaction 5.0

(Default = 5 l. May be increased up to 10 l if needed)

Adapter Stock Concentration to use 0.3

KAPA Single-Indexed Adapter Concentration to use 1.5

Dilution factor 5.0

Number of libraries per index 39

Total number of libraries with Set A + B (24 adapters) 936

Total number of libraries with Set A or B (12 adapters) 468

To confirm actual adapter:insert molar ratio:

Enter actual Adapter Stock Concentration used 15.0
Volume of Adapter used per ligation reaction 5.0
Moles of adapter used 75.0
Actual adapter:insert molar ratio 9900.0
 DNA Library Construction

cells. Refer to the Notes for additional instructions. If a cell turns red, the entered or calculated value(

ng

bp
pmol

pmol

39

936

468

M
L
pmol
DNA Library Construction

tes for additional instructions. If a cell turns red, the entered or calculated value(s) are either out of ra

Notes:

Most fields are set to round down to a single decimal, but this degree of precision is not required in pr
resolution, or to facilitate pipetting. The calculated Adapter Stock Concentration may not fully corresp
Sheet (TDS) supplied with your library preparation kit, as a result of rounding differences.

For KAPA Hyper Prep and KAPA HyperPlus Kits, refer to Table 2 in the KAPA Single-Indexed
guidelines on adapter:insert molar ratios. For the KAPA HTP and LTP Library Preparation K

With both kits, low-input and challenging samples may benefit from higher adapter:insert molar ratios
details.

If the calculated Adapter Stock Concentration exceeds 30 M, the volume of adapter added to
volume of water included in the reaction decreased proportionally. To achieve a practical solution (calc

- First, reduce the adapter:insert molar ratio slightly, as rounding my cause the stock concentratio

- If you end up having to adjust the adapter: insert molar ratio to a value that is well out of the recomm
volume of adapter used per ligation reaction (cell C16), up to a maximum of 10 L. The a
reaction has to be reduced accordingly..

- If an appropriately reduced adapter:insert molar ratio and adapter volume of 10 L does not return a
30 M or less, the input into library construction has to be reduced for that fragment length.

KAPA Single-Indexed Adapter Kits contain 40 uL of each index, which is sufficient for 8 libraries at the
contain a 10% overage, which is more than sufficient for for manual pipetting, but may not support al
methods. For such workflows, the number of libraries that can be prepared per index or kit will be low
KAPA Single-Indexed Adapter Kit Calculator:

Instructions: Select your RNA-Seq library construction worfkflow and enter your input R

Kit
Workflow

Input into RiboErase

Input into mRNA Capture

Input into library construction

Adapter Stock Concentration to use

KAPA Single-Indexed Adapter Concentration to use

Dilution factor (X)

Number of libraries per index

Total number of libraries with Set A + B (24 adapters)
Total number of libraries with Set A or B (12 adapters)
culator: Library Construction for RNA-Seq

ow and enter your input RNA amount in the appropriate yellow block.

KAPA Stranded RNA-Seq Library Preparation Kit

With RiboErase mRNA Capture Total RNA With Ribo

100 ng

100 ng

10 ng

140.0 140.0 350.0 nM

1.5 M

10.7 10.7 4.3

85 85 34
2,040 2,040 816
1,020 1,020 408
 KAPA RNA Hyper
With RiboErase mRNA Capture Total RNA

499 ng

500 ng

51

High Quality High Quality High Quality

1.5 7.0 15.0

1.5 30 30

1.0 4.3 2.0

8 34 16
192 816 384
96 408 192
ng