单细胞DNA文库制备试剂盒-文库及构建-试剂-生物在线
单细胞DNA文库制备试剂盒

单细胞DNA文库制备试剂盒

商家询价

产品名称： 单细胞DNA文库制备试剂盒

英文名称： PicoPLEX DNA-Seqkit for Illumina NGS Platforms

产品编号： R300381

产品价格： 0

产品产地： 美国

品牌商标： RUBICON GENOMICS

更新时间： null

使用范围： null

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Overview

Create more libraries with greater sensitivity!  ThruPLEX DNA-seq builds on the innovative ThruPLEX chemistry to generate DNA libraries with expanded multiplexing capability and even greater diversity with lower duplication rates. Kits contain either 48 or 96 dual read Illumina-compatible indexes pre-dispensed and sealed in barcoded microplates. Starting with as little as 50 pg of fragmented double-stranded DNA or cDNA, the entire ThruPLEX DNA-seq workflow is still performed in a single tube or well about 2 hours and requires no purification steps or sample transfers. With a low input requirement, ThruPLEX DNA-seq produces uniform GC coverage and improves molecular complexity yielding reproducible sequencing results. ThruPLEX DNA-seq can be used in any DNA-seq application, RNA-seq, ChIP-seq and offers robust target enrichment performance with all of the leading platforms.

ThruPLEX DNA-seq Kit is  protected by U.S. Patents 7,803,550; 8,071,312; 8,399,199; 8,728,737 and corresponding foreign patents. Additional patents are pending.

  • Expanded multiplexing capability – up to 96 indexes
  • Higher performance from lowest input – 50 pg to 50 ng
  • Fast and simple workflow – 3 steps in a single tube or well in about 2 hours, no transfers necessary
  • Compatible with major target enrichment platforms – Agilent SureSelect®, Roche NimbleGen® SeqCap® EZ, and IDT xGen® Lockdown® probes

 

Workflow

ThruPLEX-DNA-seq-TubeWorlflow-for-web

 

Even Better Performance at Low Input Amounts

NGS libraries were prepared from the indicated amount of DNA using either ThruPLEX DNA-seq Kit or ThruPLEX-FD Prep Kit and sequenced on a MiSeq, v3 flow cell. Picard was used to calculate estimated library size, and % duplicates at 25M read pairs. ThruPLEX DNA-seq showed greater library diversity (left) and had fewer duplicates (right) at each input amount.

ThruPLEX DNA-seq Outperforms Other NGS LIbrary Prep Kits

NGS libraries were prepared from the indicated amount of DNA using the library preparation kits indicated and sequenced on a MiSeq, v3 flow cell. (Left) Note that in every case, both ThruPLEX–FD (light blue) and ThruPLEX DNA-seq (gold) yielded more diverse libraries with less input DNA than any of the other kits. (Middle) ThruPLEX libraries had fewer duplicates at 25M read pairs than other kits. (Right) Both ThruPLEX kits have a lower and broader recommended input range than the other kits.

ThruPLEX DNA-seq Minimizes GC Bias

Relative coverage of the human genome as a function of %GC (50 ng input DNA). (Left) ThruPLEX DNA-seq has excellent coverage throughout the entire range. (Right) ThruPLEX DNA-seq (gold) performs as well as or better than other kits tested. Note that the optimal coverage should be equal to 1 in these graphs. GC composition distribution of the human genome is represented by the red bars.

More Uniform Coverage

Coverage data from the human CHR 1 at 50 ng input DNA demonstrates the superior performance of ThruPLEX DNA-seq.

 
Even Better Performance at Low Input Amounts
ThruPLEX DNA-seq Outperforms Other NGS LIbrary Prep Kits
ThruPLEX DNA-seq Minimizes GC Bias
More Uniform Coverage
ThruPLEX DNA-seq Workflow Advantage: High Quality NGS Libraries in About 2 Hours
 
 

 

Store at -20 °C.

Guaranteed for 12 months at -20°C in a constant temperature freezer.

ThruPLEX DNA-seq  Kit is designed to work on a wide variety of DNA samples with amounts varying between 50 pg and 50 ng.  To ensure that your experiment delivers quality data, the table below provides a range of recommended input amounts of  human DNA based on (1) your application and (2) the source material.  The actual amount that you chose to use may vary from those suggested below.  Note that as the amount of input DNA decreases, the diversity of the library may also decrease

Application Input sample amount (50 – 500 bp ds DNA) Recommended input amount
WGS, WES, SNV, indels, SNP, STR gDNA 10 – 50 ng
FFPE DNA 20 – 50 ng
plasma DNA 1 – 50 ng
CNV plasma, gDNA or FFPE DNA 50 pg – 50 ng*
ChIP-seq ChIP DNA 50 pg – 50 ng*
RNA-seq cDNA 50 pg – 50 ng*
     

*Use of  less than 50 pg has been cited in scientific publications.

Click here to read all FAQ’s on ThruPLEX DNA-seq Kit

1. What are the major differences between ThruPLEX®-FD Prep Kit and the new ThruPLEX DNA-seq Kit?

  • Increased multiplexing capability from 12 6nt single indexes to up to 96 8nt dual indexes (Illumina TruSeq HT) or up to 48 8nt single indexes (Sanger, see Appendix 1 of the ThruPLEX DNA-seq Kit Instruction Manual)
  • Total reaction volume lowered from 75 µL to 50 µL to accommodate more thermal cyclers
  • Protocol modified to account for reduced volume
  • Fewer PCR amplification cycles during Stage 5 of the Library Amplification Step
  • Indexing reagents for kits containing 48 or 96 indexes are pre-dispensed into a 96-well plate

2. Is ThruPLEX DNA-seq Kit compatible with any target enrichment systems?

Yes, ThruPLEX DNA-seq Kit is compatible with the major exome and target enrichment products, including Agilent SureSelect®, Roche NimbleGen® SeqCap® EZ and custom panels. ThruPLEX DNA-seq Kit target enrichment protocols and application notes can be accessed at: http://rubicongenomics.com/applications/enrichment/.

3. What DNA fragment size do you recommend?

ThruPLEX DNA-seq Kit can accommodate a range of 50 – 1,000 bp DNA input. Fragment DNA to the size specified for the Illumina platform you are using, generally between 200 and 500 bp.

4. What is the recommended elution/resuspension buffer for input DNA sample(s)? Are there any buffer concentration considerations?

The DNA sample(s) should be eluted/resuspended in a low-salt and low-EDTA, buffered solution. The preferred buffer is TE with ≤ 10 mM Tris and ≤ 0.1 mM EDTA. Avoid phosphate containing buffers.

5. Do I need to perform cleanup on my DNA sample(s) after shearing?

No, simply proceed to the Template Preparation Step (Section D.I.) in the ThruPLEX DNA-seq Kit Instruction Manual in order to preserve your total DNA and the overall diversity of the library.

6. What types of DNA samples can be used with ThruPLEX DNA-seq Kit?

  • Fragmented, double-stranded DNA, such as that from:
  • Mechanically-sheared DNA
  • Enzymatically-fragmented DNA
  • Low molecular weight cell-free DNA from plasma, urine, and other biofluids
  • Double-stranded cDNA
  • Chromatin Immunoprecipitated (ChIP) DNA
  • Sonicated DNA from formalin-fixed material (FFPE)

7. Can RNA be used with ThruPLEX DNA-seq Kit?

Yes, but RNA needs to be converted into double-stranded cDNA and fragmented to proper size prior to preparation with ThruPLEX DNA-seq Kit.

8. Can plasma, urine, or other biofluids be directly used with ThruPLEX DNA-seq Kit?

No, the template DNA must be first extracted with a DNA extraction/purification kit. Since DNA from these sources is normally degraded/fragmented, additional fragmentation is unnecessary. The volume of the sample used in the
ThruPLEX DNA-seq Kit must be no more than 10 µL.

9. Can I prepare samples for both single- and paired-end NGS sequencing?

Yes, ThruPLEX DNA-seq Kit is compatible with both single- and paired-end sequencing. The final amplified libraries will contain Illumina-compatible index sequences.

10. Is there a required DNA sample input volume for ThruPLEX DNA-seq Kit?

Yes, the required DNA sample volume is 10 µL.

ThruPLEX DNA-seq contains everything needed, including indexes in a barcoded 96-well plate.

CAT. NO. R400406 ThruPLEX DNA-seq 48D Kit (48 reactions, 48 dual indexes)

CAT. NO. R400407 ThruPLEX DNA-seq 96D Kit (96 reactions, 96 dual indexes)

CAT. NO. R400427 ThruPLEX DNA-seq 48S Kit (48 reactions, 48 single indexes)

CAT. NO. R400428 ThruPLEX DNA-seq 12S (48) Kit (48 reactions, 12 single indexes)

CAT. NO. R400429 ThruPLEX DNA-seq 12S  Kit (12 reactions, 12 single indexes)