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本篇內(nèi)容主要講解“怎么用qiime2分類器建立SILVA數(shù)據(jù)庫(kù)”,感興趣的朋友不妨來(lái)看看。本文介紹的方法操作簡(jiǎn)單快捷,實(shí)用性強(qiáng)。下面就讓小編來(lái)帶大家學(xué)習(xí)“怎么用qiime2分類器建立SILVA數(shù)據(jù)庫(kù)”吧!
利用工具建立數(shù)據(jù)庫(kù) rescript
qiime rescript get-silva-data \--p-version '138' \ --p-target 'SSURef_NR99' \ --p-include-species-labels \ --o-silva-sequences silva-138-ssu-nr99-seqs.qza \ --o-silva-taxonomy silva-138-ssu-nr99-tax.qza
這個(gè)代碼自動(dòng)獲取 99相似性的序列和分類信息,由于網(wǎng)絡(luò)原因一般運(yùn)行報(bào)錯(cuò)
wget -c https://data.qiime2.org/2020.8/common/silva-138-99-seqs.qza wget -c https://data.qiime2.org/2020.8/common/silva-138-99-tax.qza ln -s silva-138-99-tax.qza silva-138-ssu-nr99-tax.qza ln -s silva-138-99-seqs.qza silva-138-ssu-nr99-seqs.qza
#remove sequences that contain 5 or more ambiguous bases (IUPAC compliant ambiguity bases) and any homopolymers that are 8 or more bases in length qiime rescript cull-seqs \ --i-sequences silva-138-ssu-nr99-seqs.qza \ --o-clean-sequences silva-138-ssu-nr99-seqs-cleaned.qza #長(zhǎng)度過濾 qiime rescript filter-seqs-length-by-taxon \ --i-sequences silva-138-ssu-nr99-seqs-cleaned.qza \ --i-taxonomy silva-138-ssu-nr99-tax.qza \ --p-labels Archaea Bacteria Eukaryota \ --p-min-lens 900 1200 1400 \ --o-filtered-seqs silva-138-ssu-nr99-seqs-filt.qza \ --o-discarded-seqs silva-138-ssu-nr99-seqs-discard.qza #重復(fù)序列合并 qiime rescript dereplicate \ --i-sequences silva-138-ssu-nr99-seqs-filt.qza \ --i-taxa silva-138-ssu-nr99-tax.qza \ --p-rank-handles 'silva' \ --p-mode 'uniq' \ --o-dereplicated-sequences silva-138-ssu-nr99-seqs-derep-uniq.qza \ --o-dereplicated-taxa silva-138-ssu-nr99-tax-derep-uniq.qza #全長(zhǎng)分類器構(gòu)建 qiime feature-classifier fit-classifier-naive-bayes \ --i-reference-reads silva-138-ssu-nr99-seqs-derep-uniq.qza \ --i-reference-taxonomy silva-138-ssu-nr99-tax-derep-uniq.qza \ --o-classifier silva-138-ssu-nr99-classifier.qza ##特異引物分類器構(gòu)建1 #截取序列 qiime feature-classifier extract-reads \ --i-sequences silva-138-ssu-nr99-seqs-derep-uniq.qza \ --p-f-primer GTGYCAGCMGCCGCGGTAA \ --p-r-primer GGACTACNVGGGTWTCTAAT \ --p-n-jobs 2 \ --p-read-orientation 'forward' \ --o-reads silva-138-ssu-nr99-seqs-515f-806r.qza #合并重復(fù) qiime rescript dereplicate \ --i-sequences silva-138-ssu-nr99-seqs-515f-806r.qza \ --i-taxa silva-138-ssu-nr99-tax-derep-uniq.qza \ --p-rank-handles 'silva' \ --p-mode 'uniq' \ --o-dereplicated-sequences silva-138-ssu-nr99-seqs-515f-806r-uniq.qza \ --o-dereplicated-taxa silva-138-ssu-nr99-tax-515f-806r-derep-uniq.qza #構(gòu)建分類器 qiime feature-classifier fit-classifier-naive-bayes \ --i-reference-reads silva-138-ssu-nr99-seqs-515f-806r-uniq.qza \ --i-reference-taxonomy silva-138-ssu-nr99-tax-515f-806r-derep-uniq.qza \ --o-classifier silva-138-ssu-nr99-515f-806r-classifier.qza ##特異引物分類器構(gòu)建2 # 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and. 806R (5′-GGACTACHVGGGTWTCTAAT-3′) #截取序列 qiime feature-classifier extract-reads \ --i-sequences silva-138-ssu-nr99-seqs-derep-uniq.qza \ --p-f-primer ACTCCTACGGGAGGCAGCAG \ --p-r-primer GGACTACHVGGGTWTCTAAT \ --p-n-jobs 2 \ --p-read-orientation 'forward' \ --o-reads silva-138-ssu-nr99-seqs-338f-806r.qza #合并重復(fù) qiime rescript dereplicate \ --i-sequences silva-138-ssu-nr99-seqs-338f-806r.qza \ --i-taxa silva-138-ssu-nr99-tax-derep-uniq.qza \ --p-rank-handles 'silva' \ --p-mode 'uniq' \ --o-dereplicated-sequences silva-138-ssu-nr99-seqs-338f-806r-uniq.qza \ --o-dereplicated-taxa silva-138-ssu-nr99-tax-338f-806r-derep-uniq.qza #構(gòu)建分類器 qiime feature-classifier fit-classifier-naive-bayes \ --i-reference-reads silva-138-ssu-nr99-seqs-338f-806r-uniq.qza \ --i-reference-taxonomy silva-138-ssu-nr99-tax-338f-806r-derep-uniq.qza \ --o-classifier silva-138-ssu-nr99-338f-806r-classifier.qza
注意:qiime2建立分類數(shù)據(jù)庫(kù)很消耗內(nèi)存,至少50G以上
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