TABLE 1.
Primers, gRNA, and probe used to establish the K. pneumoniae CRISPR-top assay
| Primer | Type | Sequence (5′–3′)a | Lengthb |
|---|---|---|---|
| Kp1-F3 | Forward outer primer | GCTGTTTGTTATCTTTATGTCG | 22 nt |
| Kp1-B3 | Backward outer primer | CATCCACATTTGCAGCATAT | 20 nt |
| Kp1-FIP | Forward inner primer | TGGGGTTATCGATTTTGAACTGATT-TTC-GGCGAACATCCATTTTGAC | 47 nt |
| Kp1-BIP | Backward inner primer | ACAAAAACACCAGTGTAGGGC-GATTCTGTTTTACTCAGTGACAAT | 45 nt |
| Kp1-LF | Loop forward primer | CAGATTCTTCCGTACCCGC | 19 nt |
| gRNA | Guide RNA | GUCUAGAGGACAGAAUUUUUCAACGGGUGUGCCAAUGGCCACUUUCCAGGUGGCAAAGCCCGUUGAGCUUCUCAAAUCUGAGAAGUGGCACGGCGAACAUCCAUUUUGACC | 111 nt |
| Probe | SsDNA fluorescence probe | FAM-TTATTATTAT-BHQ1 | 10 nt |
a
The inserted PAM site (TTC) is in bold.
bnt, nucleotide.