Linking sequence reads using paired code tags
Artificial transposon sequences having code tags and target nucleic acids containing such sequences. Methods for making artificial transposons and for using their properties to analyze target nucleic acids.
1. An artificial transposon, comprising:
a first transposase recognition site,
a second transposase recognition site,
a barcode disposed therebetween, wherein the barcode comprises a double-stranded nucleic acid sequence comprising
a first strand barcode and
a second strand barcode, wherein the first strand barcode and the second strand barcode comprise non-complementary sequences; and
a fragmentation site disposed within the first strand barcode and the second strand barcode.
2. The transposon of claim 1 , wherein the first transposase recognition site comprises a hyperactive Tn5 transposase recognition site.
3. The transposon of claim 1 , wherein the first transposase recognition site comprises a Mu transposase recognition site.
4. The transposon of claim 1 , wherein the first transposase recognition site comprises an IS911 transposase recognition site.
5. The transposon of claim 1 , further comprising at least one universal primer site.
6. An isolated template nucleic acid, comprising at least a portion of a target nucleic acid or copy thereof and at least a first transposon and second transposon of claim 1 , wherein the barcode of the first transposon is different from the barcode of the second transposon.
7. The transposon of claim 1 , wherein both the first strand barcode and the second strand barcode are at least four nucleotides in length.
8. The transposon of claim 1 , wherein the fragmentation site comprises a nickase recognition site.
9. A DNA library preparation kit, comprising:
a plurality of artificial transposons, wherein each artificial transposon of the plurality of artificial transposon comprises:
a first transposase recognition site;
a second transposase recognition site;
a barcode disposed therebetween, wherein the barcode comprises a double-stranded nucleic acid sequence comprising
a first strand barcode and
a second strand barcode, wherein the first strand barcode and the second strand barcode comprise non-complementary sequences; and
a fragmentation site disposed within the first strand barcode and the second strand barcode; and
wherein each barcode of each artificial transposon of the plurality of artificial transposons is distinguishable from one another.
10. The DNA library preparation kit of claim 9 , wherein both the first strand barcode and the second strand barcode are at least four nucleotides in length.