HLA-C*07 Product Insert Page 1 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU) Lot No.: 7E6 Lot-specific Information April 2019 For In Vitro Diagnostic Use Rev. No.: 04 Olerup SSP ® HLA-C*07 Product number: 101.615-24/04 – including Taq pol. 101.615-24u/04u – without Taq pol. Lot number: 7E6 Expiry date: 2021-03-01 Number of tests: 24 tests – Product No. 101.615-24/24u 4 tests – Product No. 101.615-04/04u Number of wells per test: 93+1 Storage - pre-aliquoted primers: dark at -20 o C - PCR Master Mix: -20 o C - Adhesive PCR seals RT - Product Insert RT This Product Description is only valid for Lot No. 7E6. Complete product documentation consists of generic Instructions for Use (IFU), lot specific Product Insert, Worksheet and Certificate. CHANGES COMPARED TO THE PREVIOUS OLERUP SSP ® HLA-C*07 LOT (4D7) The HLA-C*07 kit is updated for new alleles to enable separation of: • Confirmed 1 alleles as listed in the IMGT/HLA database • Polymorphisms in exons outside of the region encoding the peptide binding domain • Null and Alternatively expressed alleles 1 As described in section Uniquely Identified Alleles. The HLA-C*07 specificity and interpretation tables have been updated for the HLA- C alleles described since the previous Olerup SSP ® HLA-C*07 lot was made (Lot No. 4D7). The kit design is based on IMGT/HLA database 3.26.0. As of lot series V, the Specificity Table is included in the lot-specific Product Insert, and the Interpretation Table is included in the Worksheet.
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Olerup SSP HLA-C*07 - CareDx · 2019. 6. 18. · C alleles described since the previous Olerup SSP® HLA-C*07 lot was made (Lot No. 4D7). The kit design is based on IMGT/HLA database
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HLA-C*07 Product Insert Page 1 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
Olerup SSP® HLA-C*07 Product number: 101.615-24/04 – including Taq pol. 101.615-24u/04u – without Taq pol. Lot number: 7E6 Expiry date: 2021-03-01 Number of tests: 24 tests – Product No. 101.615-24/24u 4 tests – Product No. 101.615-04/04u Number of wells per test: 93+1 Storage - pre-aliquoted primers: dark at -20oC - PCR Master Mix: -20oC - Adhesive PCR seals RT - Product Insert RT
This Product Description is only valid for Lot No. 7E6. Complete product documentation consists of generic Instructions for Use (IFU), lot
specific Product Insert, Worksheet and Certificate.
CHANGES COMPARED TO THE PREVIOUS OLERUP SSP®
HLA-C*07 LOT (4D7)
The HLA-C*07 kit is updated for new alleles to enable separation of:
• Confirmed1 alleles as listed in the IMGT/HLA database
• Polymorphisms in exons outside of the region encoding the peptide binding domain
• Null and Alternatively expressed alleles
1As described in section Uniquely Identified Alleles.
The HLA-C*07 specificity and interpretation tables have been updated for the HLA-C alleles described since the previous Olerup SSP® HLA-C*07 lot was made (Lot
No. 4D7). The kit design is based on IMGT/HLA database 3.26.0. As of lot series V, the Specificity Table is included in the lot-specific Product Insert, and the Interpretation Table is included in the Worksheet.
HLA-C*07 Product Insert Page 2 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
The primers of the wells detailed below have been exchanged, added or modified compared to the previous lot. Well 5’-primer 3’-primer rationale
18 - Added 3’-primer added for the C*07:486 allele. 27 Added - 5’-primer added for the C*07:322 allele. 28 Added - 5’-primer added for the C*07:396 allele. 30 Modified - 5’-primer modified for improved HLA-specific
amplification of the C*07:75 and C*07:254 alleles. 31 - Added,
removed 3’-primer added for the C*07:276 allele, 3’-primer removed for improved HLA-specific amplification.
34 - Modified 3’-primer modified for improved HLA-specific amplification.
38 Exchanged - 5’-primer exchanged for decreased tendency of primer oligomer formation.
42 Added - 5’-primer added for the C*07:352 allele. 43 - Added 3’-primer added for the C*07:445 allele. 44 Added - 5’-primer added for the C*07:396 allele. 48 - Added 3’-primer added for the C*07:276 allele. 49 Added - 5’-primer added for the C*07:330:02 allele. 50 - Added 3’-primer added for the C*07:310 allele. 51 - Added 3’-primer added for the C*07:483N allele. 53 Added - 5’-primer added for the C*07:330:02 allele. 54 - Added 3’-primer added for the C*07:547 allele. 56 Added - 5’-primer added for the C*07:524 allele. 57 Added - 5’-primer added for the C*07:524 allele. 59 Added - 5’-primer added for the C*07:485 allele. 60 Added - 5’-primer added for the C*07:485 allele. 61 Removed Removed,
added 3’- and 5’-primers removed for improved HLA-specific amplification, 3’-primers added for the C*07:450 and C*07:762 alleles.
62 Added - 5’-primer added for the C*07:322 allele. 63 Added - 5’-primer added for the C*07:425 allele. 64 Added - 5’-primer added for the C*07:326 allele. 65 - Added 3’-primers added for the C*07:494Q and
C*07:551N alleles. 66 - Added 3’-primer added for the C*07:513Q allele. 67 - Added 3’-primer added for the C*07:456 allele. 68 - Added 3’-primer added for the C*07:456 allele. 69 - Added 3’-primer added for the C*07:494Q allele. 70 - Added 3’-primer added for the C*07:310 allele. 71 Added - 5’-primer added for the C*07:425 allele. 72 Added - 5’-primer added for the C*07:484N allele. 73 - Added 3’-primer added for the C*07:513Q allele. 75 Added Added Primer pairs added for the C*07:533 and
C*07:544 alleles. 77 - Added 3’-primer added for the C*07:547 allele. 80 - Added 3’-primer added for the C*07:486 allele. 82 - Added 3’-primer added for the C*07:483N allele.
HLA-C*07 Product Insert Page 3 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
83 - Added 3’-primers added for the C*07:445 and C*07:476N alleles.
84 - Added 3’-primers added for the C*07:439 and C*07:551N alleles.
85 Added - 5’-primer added for the C*07:360 allele. 86 Added - 5’-primer added for the C*07:226 allele. 87 - Added 3’-primers added for the C*07:411 and C*07:507N
alleles. 88 Added Added Primer pairs added for the C*07:491:01N,
C*07:491:02N and C*07:507N alleles. 89 Added - 5’-primer added for the C*07:326 allele. 92 - Added 3’-primer added for the C*07:457 allele. 93 Added Added Primer pairs added for the C*07:491:01N and
C*07:491:02N alleles.
Change in revision R01 compared to R00: 1. Primer mix 80 does not amplifies the C*07:316 and 07:359 and the C*05:108 and C*06:111 alleles. Thus, this lot of the C*07 subtyping kit cannot distinguish the C*07:359 and the C*07:02:01:01-07:02:77, 07:105, 07:114, 07:135, 07:144, 07:146, 07:157-07:160, 07:167, 07:169, 07:174-07:175, 07:178, 07:185, 07:187, 07:193, 07:195, 07:208-07:209, 07:211, 07:213, 07:217, 07:221, 07:233, 07:239-07:240, 07:251, 07:261-07:262, 07:270-07:271, 07:273-07:275, 07:283-07:284, 07:286-07:287, 07:290-07:291, 07:305-07:306, 07:309, 07:312, 07:318-07:321, 07:325, 07:327, 07:333-07:334, 07:341:01-07:341:02, 07:344-07:346, 07:363, 07:372, 07:376, 07:379, 07:382-07:384, 07:388-07:389, 07:391-07:392, 07:398-07:401, 07:405, 07:413-07:416, 07:432-07:434, 07:436, 07:438, 07:449, 07:464-07:465, 07:472-07:473, 07:477-07:478, 07:482, 07:489, 07:495-07:496, 07:504-07:505, 07:509-07:511, 07:516, 07:518, 07:520, 07:525, 07:529-07:530, 07:532, 07:539, 07:541-07:542, 07:548, 07:553 and 07:560 alleles. This has been corrected in the Specificity and Interpretation Tables. Change in revision R02 compared to R01: 1. Primer mix 75 does not amplifies the C*07:533 and the C*15:97 alleles. Thus, this lot of the C*07 subtyping kit cannot distinguish the C*07:533 and the C*07:02:01:01-07:02:77, 07:105, 07:114, 07:135, 07:144, 07:146, 07:157-07:160, 07:167, 07:169, 07:174-07:175, 07:178, 07:185, 07:187, 07:193, 07:195, 07:208-07:209, 07:211, 07:213, 07:217, 07:221, 07:233, 07:239-07:240, 07:251, 07:261-07:262, 07:270-07:271, 07:273-07:275, 07:283-07:284, 07:286-07:287, 07:290-07:291, 07:305-07:306, 07:309, 07:312, 07:318-07:321, 07:325, 07:327, 07:333-07:334, 07:341:01-07:341:02, 07:344-07:346, 07:359, 07:363, 07:372, 07:376, 07:379, 07:382-07:384, 07:388-07:389, 07:391-07:392, 07:398-07:401, 07:405, 07:413-07:416, 07:432-07:434, 07:436, 07:438, 07:449, 07:464-07:465, 07:472-07:473, 07:477-07:478, 07:482, 07:489, 07:495-07:496, 07:504-07:505, 07:509-07:511, 07:516, 07:518, 07:520, 07:525, 07:529-07:530, 07:532, 07:539, 07:541-07:542, 07:548, 07:553 and 07:560 alleles. This has been corrected in the Specificity and Interpretation Tables. Change in revision R03 compared to R02: 1. Primer mix 78 does not amplifies the C*07:68, 07:76:02 and the C*06:136 alleles. This has been corrected in the Specificity and Interpretation Tables. Change in revision R04 compared to R03: 1. The expiration date has been altered due to extension of shelf-life.
HLA-C*07 Product Insert Page 4 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
Well 94 contains Negative Control primer pairs, that will amplify more than 95% of the Olerup SSP® HLA Class I, DRB, DQB1, DPB1 and DQA1 amplicons as well as all the amplicons generated by the control primer pairs matching the human growth hormone gene.
HLA-specific PCR product sizes range from 75 to 200 base pairs. The PCR product generated by the positive control primer pair is 430 base pairs.
1The nucleotide position for HLA class I genes and the codon for HLA class II genes, in the 2nd or 3rd exon, matching the specificity-determining 3'-end of the primer is given. Nucleotide and codonnumbering as on the www.ebi.ac.uk/imgt/hla web site. The sequence of the 3 terminal nucleotides of the primer is given. 2The nucleotide position for HLA class I genes and the codon for HLA class II genes, in the 2nd or 3rd exon or the 2nd intron, matching the specificity-determining 3'-end of the primer is given in the anti-sense direction. Nucleotide and codon numbering as on the www.ebi.ac.uk/imgt/hla web site. The sequence of the 3 terminal nucleotides of the primer is given.
HLA-C*07 Product Insert Page 5 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
PRODUCT DESCRIPTION
HLA-C*07 SSP subtyping CONTENT The primer set contains 5’- and 3’-primers for identifying the HLA-C*07:01 to HLA-C*07:563 alleles.
PLATE LAYOUT Each test consists of 94 PCR reactions in a 96 well cut PCR plate. Wells 95 to 96 are empty.
1 2 3 4 5 6 7 8
9 10 11 12 13 14 15 16
17 18 19 20 21 22 23 24
25 26 27 28 29 30 31 32
33 34 35 36 37 38 39 40
41 42 43 44 45 46 47 48
49 50 51 52 53 54 55 56
57 58 59 60 61 62 63 64
65 66 67 68 69 70 71 72
73 74 75 76 77 78 79 80
81 82 83 84 85 86 87 88
89 90 91 92 93 NC empty empty
The 96 well cut PCR plate is marked with ‘HLA-C*07’ in silver/gray ink. Well No. 1 is marked with the Lot No. ‘7E6’. Wells 1 to 93 – HLA-C*07 high resolution primers. Well 94 – Negative Control (NC). A faint row of numbers is seen between wells 1 and 2 or wells 7 and 8 of the PCR trays. These stem from the manufacture of the trays, and should be disregarded. The PCR plates are heat-sealed with a PCR-compatible foil.
INTERPRETATION Due to the sharing of sequence motifs between HLA-C alleles, non-HLA-C*07 alleles will be amplified by primer mixes 1 to 3, 6, 8 to 14, 16 to 20, 22, 25 to 27, 30, 32, 36 to 38, 40, 42, 45, 49, 52, 53, 55, 57, 58, 60 to 62, 64 to 66, 68, 69, 71 to 73, 75, 77 to 81, 84, 88 to 90, 92 and 93. In addition, a few HLA-A and HLA-B alleles will be amplified by primer mixes 2, 4, 6 to 8, 10 to 12, 14, 16, 18, 20, 22, 24, 25, 27, 32, 37, 38, 41, 42, 45, 53 to 56, 58 to 62, 65, 66, 69, 71 to 73, 77, 79, 81, 84, 88, 90, 92 and 93. For further details see Specificity Table.
HLA-C*07 Product Insert Page 6 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
UNIQUELY IDENTIFIED ALLELES All the HLA-C*07 alleles, i.e. C*07:01 to C*07:563, recognized by the HLA Nomenclature Committee in October 20161,2 will be amplified by the primers in the HLA-C*07 SSP kit3,4. The HLA-C*07 kit enables separation of the confirmed HLA-C*07 alleles as listed in the IMGT/HLA database. An HLA allele is listed as confirmed by IMGT/HLA if it has been sequenced by more than a single laboratory or from multiple sources. Current allele confirmation status for HLA-C*07 alleles is listed below. The HLA-C*07 kit also enables identification of polymorphisms in exons outside of the region encoding the peptide binding domain and of null and alternatively expressed alleles. The following HLA-C*07 alleles can be distinguished by the different sizes of the HLA-specific PCR product: Alleles Primer mix Alleles Primer mix
The HLA-C*07 subtyping kit cannot distinguish the following silent mutations: the C*07:01:01:01-07:01:01:07, 07:01:03-07:01:04, 07:01:06-07:01:12, 07:01:14-07:01:24, 07:01:26, 07:01:28-07:01:35, 07:01:37-07:01:51, 07:01:53-07:01:58, 07:204:01-07:204:02 and 07:454:01 alleles, the 07:01:02, 07:01:27 and 07:01:52 alleles, the C*07:01:05 and 07:01:36 alleles, the C*07:02:01:01-07:02:77 and 07:341:01-07:341:02 alleles, the C*07:04:01:01-07:04:11 alleles, the C*07:17:01-07:17:03 alleles, the 07:26:01-07:26:02 alleles, the C*07:27:01-07:27:02 alleles, the C*07:29:01-07:29:02 alleles, the C*07:31:01-07:31:02 alleles, the 07:38:01-07:38:02 alleles, the 07:43:01-07:43:02 alleles, the C*07:56:01-07:56:02, the C*07:78:01-07:78:02 alleles, the C*07:96:01-07:96:02 alleles, the C*07:108:01-07:108:02 alleles, the C*07:131:01-07:131:02 alleles, the C*07:137:01-07:137:02 alleles, the C*07:141:01-07:141:02 alleles, the C*07:172:01-07:172:02 alleles, the C*07:199:01-07:199:02 alleles, the 07:260:01-07:260:02 alleles, the C*07:314:01-07:314:02 alleles, the 07:368:01-07:368:02 or the 07:491:01N-07:491:02N alleles.
HLA-C*07 Product Insert Page 7 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
1HLA-C alleles listed on the IMGT/HLA web page 2016-October-14, release 3.26.0, www.ebi.ac.uk/imgt/hla. 2Alleles that have been deleted from or renamed in the official WHO HLA Nomenclature up to and including the last IMGT/HLA database release can be retrieved from web page http://hla.alleles.org/alleles/deleted.html. 3The C*07:27:01-07:27:02 and 07:556 and the C*12:181 alleles will give rise to identical amplification patterns. These alleles can e.g. be distinguished by the HLA-C low resolution kit. 4This lot of the C*07 subtyping kit cannot distinguish the C*07:359 and the C*07:02:01:01-07:02:77, 07:105, 07:114, 07:135, 07:144, 07:146, 07:157-07:160, 07:167, 07:169, 07:174-07:175, 07:178, 07:185, 07:187, 07:193, 07:195, 07:208-07:209, 07:211, 07:213, 07:217, 07:221, 07:233, 07:239-07:240, 07:251, 07:261-07:262, 07:270-07:271, 07:273-07:275, 07:283-07:284, 07:286-07:287, 07:290-07:291, 07:305-07:306, 07:309, 07:312, 07:318-07:321, 07:325, 07:327, 07:333-07:334, 07:341:01-07:341:02, 07:344-07:346, 07:363, 07:372, 07:376, 07:379, 07:382-07:384, 07:388-07:389, 07:391-07:392, 07:398-07:401, 07:405, 07:413-07:416, 07:432-07:434, 07:436, 07:438, 07:449, 07:464-07:465, 07:472-07:473, 07:477-07:478, 07:482, 07:489, 07:495-07:496, 07:504-07:505, 07:509-07:511, 07:516, 07:518, 07:520, 07:525, 07:529-07:530, 07:532, 07:539, 07:541-07:542, 07:548, 07:553 and 07:560 alleles. This lot of the C*07 subtyping kit cannot distinguish the C*07:533 and the C*07:02:01:01-07:02:77, 07:105, 07:114, 07:135, 07:144, 07:146, 07:157-07:160, 07:167, 07:169, 07:174-07:175, 07:178, 07:185, 07:187, 07:193, 07:195, 07:208-07:209, 07:211, 07:213, 07:217, 07:221, 07:233, 07:239-07:240, 07:251, 07:261-07:262, 07:270-07:271, 07:273-07:275, 07:283-07:284, 07:286-07:287, 07:290-07:291, 07:305-07:306, 07:309, 07:312, 07:318-07:321, 07:325, 07:327, 07:333-07:334, 07:341:01-07:341:02, 07:344-07:346, 07:359, 07:363, 07:372, 07:376, 07:379, 07:382-07:384, 07:388-07:389, 07:391-07:392, 07:398-07:401, 07:405, 07:413-07:416, 07:432-07:434, 07:436, 07:438, 07:449, 07:464-07:465, 07:472-07:473, 07:477-07:478, 07:482, 07:489, 07:495-07:496, 07:504-07:505, 07:509-07:511, 07:516, 07:518, 07:520, 07:525, 07:529-07:530, 07:532, 07:539, 07:541-07:542, 07:548, 07:553 and 07:560 alleles.
HLA-C*07 Product Insert Page 22 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
*02:99, B*07:87, B*27:55, B*27:149
93 155 bp 200 bp
1070 bp *07:491:01N-07:491:02N *07:453
*01:111N, 14:47N, B*15:209N, B*46:55N
949 - - Negative Control
1Alleles are assigned by the presence of specific PCR product(s). However, the sizes of the specific PCR products may be helpful in the interpretation of HLA-C*07 SSP typings. When the primers in a primer mix can give rise to HLA-specific PCR products of more than one length this is indicated if the size difference is more than 20 base pairs. Size differences of 20 base pairs or less are not given. For high resolution SSP kits, the alleles listed are specified according to amplicon length. Nonspecific amplifications, i.e. a ladder or a smear of bands, may sometimes be seen. GC-rich primers have a higher tendency of giving rise to nonspecific amplifications than other primers. PCR fragments longer than the control bands may sometimes be observed. Such bands should be disregarded and do not influence the interpretation of the SSP typings. PCR fragments migrating faster than the control bands, but slower than a 400 bp fragment may be seen in some gel read-outs. Such bands can be disregarded and do not influence the interpretation of the SSP typings. Some primers may give rise to primer oligomer artifacts. Sometimes this phenomenon is an inherit feature of the primer pair(s) of a primer mix. More often it is due to other factors such as too low amount of DNA in the PCR reactions, taking too long time in setting up the PCR reactions, working at elevated room temperature or using thermal cyclers that are not pre-heated.
2The internal positive control primer pairs amplify segments of the human growth hormone gene. The internal positive control bands are 1070 or 800 base pairs respectively, well distribution as outlined in the table. Well number 1 contains the shorter, 800 bp, internal positive control band. The well distribution of the internal controls can help in orientation of the kit on gel photo, as well as allow for kit identification. In the presence of a specific amplification the intensity of the control band often decreases.
3For several HLA Class I alleles 1st and/or 4th exon(s) and beyond, as well as intron nucleotide sequences, are not available. In these instances it is not known whether some of the primers of the SSP sets are completely matched with the target sequences or not. Assumption is made that unknown sequences in these regions are conserved within allelic groups.
4Due to the sharing of sequence motifs between HLA-C alleles, non-HLA-C*07 alleles will be amplified by primer mixes 1 to 3, 6, 8 to 14, 16 to 20, 22, 25 to 27, 30, 32, 36 to 38, 40, 42, 45, 49, 52, 53, 55, 57, 58, 60 to 62, 64 to 66, 68, 69, 71 to 73, 75, 77 to 81, 84, 88 to 90, 92 and 93. In addition, a few HLA-A and HLA-B alleles will be amplified by primer mixes 2, 4, 6 to 8, 10 to 12, 14, 16, 18, 20, 22, 24, 25, 27, 32, 37, 38, 41, 42, 45, 53 to 56, 58 to 62, 65, 66, 69, 71 to 73, 77, 79, 81, 84, 88, 90, 92 and 93.
5HLA-specific PCR products shorter than 125 base pairs have a lower intensity and are less sharp than longer PCR products.
6Primer mixes 19, 20, 34, 36, 43, 44, 46, 47, 59 and 82 have a tendency to giving rise to primer oligomer formation. 7Primer mixes 13, 14, 17, 20, 23, 24, 30, 37, 40, 45, 65, 66, 70, 74, 77, 85 and 91 may have tendencies of unspecific
amplifications. 8Primer mixes 14, 15 and 16 may give rise to a lower yield of HLA-specific PCR product than the other C*07 primer
mixes. 9Primer mix 94 contains a negative control, which will amplify more than 95% of HLA amplicons as well as the
amplicons generated by the control primer pairs matching the human growth hormone gene. HLA-specific PCR product sizes range from 75 to 200 base pairs and the PCR product generated by the HGH positive control primer pair is 430 base pairs. ‘w’, might be weakly amplified.
HLA-C*07 Product Insert Page 27 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
1The internal positive control primer pairs amplify segments of the human growth hormone gene. The internal positive control bands are 1070 or 800 base pairs respectively, well distribution as outlined in the table. Well number 1 contains the shorter, 800 bp, internal positive control band. The well distribution of the internal controls can help in orientation of the kit on gel photo, as well as allow for kit identification. In the presence of a specific amplification the intensity of the control band often decreases.
2The nucleotide position matching the specificity-determining 3'-end of the primer is given. Nucleotide numbering as on the www.ebi.ac.uk/imgt/hla web site. The sequence of the 3 terminal nucleotides of the primer is given.
3The nucleotide position matching the specificity-determining 3'-end of the primer is given in the anti-sense direction. Nucleotide numbering as on the www.ebi.ac.uk/imgt/hla web site. The sequence of the 3 terminal nucleotides of the primer is given.
HLA-C*07 Product Insert Page 34 of 36 101.615-24/04 – including Taq pol., IFU-01 Visit www.labproducts.caredx.com for 101.615-24u/04u – without Taq pol., IFU-02 “Instructions for Use” (IFU)
Lot No.: 7E6 Lot-specific Information
April 2019 For In Vitro Diagnostic Use Rev. No.: 04
1The provided cell line HLA specificities are retrieved from the http://www.ihwg.org/hla web site. The specificity of an individual cell line may thus be subject to change.
2The specificity of each primer solution in the kit has been tested against 48 well characterized cell line DNAs and where applicable, additional cell line DNAs. No DNAs carrying the alleles to be amplified by primer solutions 5, 10, 11, 13, 15, 16, 18, 19, 23 to 31, 34 to 36, 38, 39, 41 to 44, 46 to 76 and 78 to 93 were available. The specificities of the primers in primer solutions 5, 10, 11, 13, 15, 16, 18, 19, 24 to 27, 31, 36, 42 to 44, 47 to 50, 53 to 60, 63, 64, 68, 71, 72, 74, 78, 79, 81, 85, and 89 to 91 were tested by separately adding one or more 5’-primers, respectively one or more 3’-primers. In primer solutions 35, 38, 46, 51, 61, 65 to 67, 69, 70, 73, 75, 80, 82 to 84, 87, 88, 92 and 93 it was only possible to test the 5’-primers, the 3’-primers were not possible to test. In primer solutions 23, 28 to 30, 34, 39, 41, 52, 62, 76 and 86 it was only possible to test the 3’-primers, the 5’-primers were not possible to test. In primer solutions 3, 8, 11, 20, 22, 26, 27, 33, 42, 44, 47 to 49, 53, 55 to 57, 59, 60, 63, 64, 71, 72, 85 and 89 one or more 5’-primers were not possible to test, and in primer solutions 4, 5, 9, 13 to 15, 18, 21, 25, 31, 32, 37, 40, 43, 47, 48, 50, 54, 58, 60, 63, 64, 68, 74, 77 and 85 one or more 3’-primers were not possible to test. Additional primers in primer solutions 2, 8, 20, 21, 33, 37, 40 and 45 were tested by separately adding one 5’-primer and/or one or two 3’-primers.