- 询价
- ZYMO RESEARCH
- 美国
- D5450
- 2026年01月05日
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- 详细信息
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- 文献和实验
- 技术资料
- 保存条件：
-70C, RT
- 英文名：
RRHP 5-hmC Library Prep Kit
- 供应商：
简石生物
HIGHLIGHTS
- Innovative library preparation for strand-specific mapping of 5-hmC in DNA.
- Streamlined workflow accommodates low (≥100ng) DNA inputs.
- Libraries are ready for Next-Generation sequencing (Illumina-compatible).
DESCRIPTION
The RRHP 5-hmC Library Prep Kit is an all-inclusive solution for analysis of genome-wide 5-hydroxymethylcytosine (5-hmC) positions at single-base resolution. The Reduced Representation Hydroxymethylation Profiling (RRHP) method is based on blocking MspI digestion by glucosylating 5-hmC within MspI recognition sites. Fragments lacking glucosylated 5-hmC at the adapter-ligation junction will be cleaved and not amplified by PCR. Therefore, only fragments containing 5-hmC will be successfully amplified and analyzed by Next-Generation Sequencing. Fragments with higher 5-hmC levels will be correlated with higher frequency of sequencing reads. RRHP bypasses the need for bisulfite conversion, which allows for DNA inputs as low as 100 ng, lower sequencing depth, and straight-forward bioinformatics processing.
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文献和实验Screening lambda cDNA or genomic libraries
MediaNeed LB plates (9 cm or 14 cm)to plate the library and do secondary screens.I typically screen about 500,000-1 million plaques for a primary cDNA screen.This fits on 11-22 14 cm plates.I do the secondary screens on 9 cm plates- you should pour
Raf 蛋白 Ras 结合结构域的简并进化库合成以及利用片段互补法快速筛选二氢叶酸还原酶的快速折叠且稳定的克隆
固体培养基。 ( 9 ) 每文库 100〜250 ml LB 培养基。LB 培养基中添加 0.2% 葡萄糖、0.25 倍的 M9 盐溶液、10 μg/ml 四环素、10 μg/ml 壮观霉素和 10 μg/ml 氨苄青霉素。 ( 10 ) Plasmid Midi Kit (12143) (Qiagen)。 2.3 文库的筛选 ( 1 ) BL21 电转感受态细胞，先转化质粒 pREP4，再转化 pQE32△F（F [ 3 ] 代表 DHFR 片段
Digital Gene Expression by Tag Sequencing on the Illumina Genome Analyzer
DEPC‐treated H 2 O ( appendix 2D ) 10 µM GEX Adapter 1 (from Illumina Tag Profiling Sample Prep Kit, cat. no. FC‐102‐1005
