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Bio-Toolkit

Reverse Complement

Generate reverse complement of DNA sequence

5'3'

Tool Guide

Definition

Reverse complement generator — Computes the complementary strand (A↔T, G↔C) of an input DNA sequence and reverses it (5'→3') to give the reverse complement. Supports IUPAC ambiguity codes (R/Y/S/W/K/M/B/D/H/V/N).

Purpose

(1) Reverse primer design — reverse complement of forward sequence = reverse primer candidate (2) Confirm antisense strand for cloning (e.g., shRNA stem-loop) (3) Check antisense strand during NGS read mapping (4) View gRNA spacer in both sense/antisense (CRISPR design)

How to Use

① Enter DNA sequence (case-insensitive, spaces/newlines auto-removed) ② Auto-displays: • Complement (same direction) • Reverse (reversed direction) • Reverse Complement (most commonly used) • Length and GC% ③ Copy button for clipboard Non-DNA characters trigger a warning. IUPAC codes (N, R, Y, etc.) convert correctly.

Examples

Example 1) Forward primer 5'-ATGGCCAAGGTGGAC-3' → Reverse complement: 5'-GTCCACCTTGGCCAT-3' → Use as reverse primer (verify matching Tm via GC/Tm tool) Example 2) gRNA target 5'-GAGTCCGAGCAGAAGAAGAA-3' (followed by PAM NGG) → Reverse complement: 5'-TTCTTCTTCTGCTCGGACTC-3' → Useful for checking PAM on antisense strand

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🔗 Bio Resources

🔬NCBI PubMed🧪AlphaFold DB🏥ClinicalTrials.gov📄bioRxiv🚀ASGCT🏛️Broad Institute

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